AU2021256235A1 - Imidazolidinone derivatives and medical use thereof - Google Patents

Imidazolidinone derivatives and medical use thereof Download PDF

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AU2021256235A1
AU2021256235A1 AU2021256235A AU2021256235A AU2021256235A1 AU 2021256235 A1 AU2021256235 A1 AU 2021256235A1 AU 2021256235 A AU2021256235 A AU 2021256235A AU 2021256235 A AU2021256235 A AU 2021256235A AU 2021256235 A1 AU2021256235 A1 AU 2021256235A1
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alkyl
halogen
compound
methyl
mmol
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AU2021256235B2 (en
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Hongzhu CHU
Lvxue HE
Yang He
Feiquan LEI
Bing Liu
Guizhuan SU
Yi Sun
Meiwei WANG
Yonggang Wei
Xuezhen XU
Jie Yan
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Chengdu Baiyu Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52—Purines, e.g. adenine
    • A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • C07D473/26—Heterocyclic compounds containing purine ring systems with an oxygen, sulphur, or nitrogen atom directly attached in position 2 or 6, but not in both
    • C07D473/32—Nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled

Abstract

The present application relates to an imidazolinone derivative and the use thereof in medicine, and specifically relates to a pyrimidine derivative as represented by general formula (I), or a stereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt or co-crystal thereof, a pharmaceutical composition containing same and the use of the compound or the composition of the present application in the preparation of drugs for treating cancers.

Description

Imidazolidinone Derivatives and Medical Use thereof
Technical Field
The present invention relates to imidazolidinone derivatives represented by general formula (I), or stereoisomers, solvates, prodrugs, metabolites, pharmaceutically acceptable salts or co-crystals thereof, a pharmaceutical composition comprising the same, and use thereof as a DNA-PK inhibitor.
Background Art
The DNA-dependent protein kinase (DNAPK) is a DNA-PK enzyme complex composed of a Ku70/Ku8O heterodimer and a DNA-dependent protein kinase catalytic subunit (DNA-PKcs). This enzyme complex requires the presence of DNA to be activated and perform the corresponding functions (George et al., 2019). As a serine/threonine protein kinase, DNA-PK is a member of the PIKK (phosphatidylinositol 3-kinase-related kinase) family, which not only plays an important role in repairing intracellular DNA double-strand breaks (DSBs) and cellular DNA recombination or antibody DNA rearrangement (V(D)J recombination), but also participates in chromosomal modification, transcription regulation, telomere maintenance, and other physiological processes.
In normal physiological processes, a variety of factors may cause DSBs in DNA. For example, DSBs often appear as intermediate products in the process of DNA recombination in somatic cells, which is a physiological process very important for the establishment of a functional immune system in all vertebrates. Furthermore, single- or double-strand breaks may be caused by damaged bases found at replication forks during DNA replication. DNA may also generate DSBs due to the attack by reactive oxygen species (ROS) during normal metabolism (Cannan & Pederson, 2016). In addition, various exogenous factors may also lead to DSBs, such as ionizing radiation (IR) and chemotherapeutic agents (such as topoisomerase II inhibitors) (George et al., 2019). If DSBs are not repaired or are incorrectly repaired, mutations and/or chromosomal aberrations will occur, eventually leading to cell death. In order to cope with the harm inflicted by DSBs, eukaryotic cells have evolved multiple mechanisms to repair damaged DNA to maintain cell viability and genome stability. In eukaryotic cells, the most important manner of DNA repair is non-homologous end-joining (NHEJ), which directly rejoins broken DNAs, does not require involvement of homologous DNA fragments, and can occur at any stage of the cell cycle. NHEJ is a dynamic process mediated by DNA-PK that requires participation of multiple proteins and signaling pathways, which basically comprises the following procedures: (1) the Ku70/Ku8O heterodimer recognizes and binds to the broken ends of double-stranded DNAs; (2) proteins such as DNA-PKcs and XRCC4-DNA ligase IV complex are recruited to both sides of the broken DNA double strands; (3) DNA-PKcs is autophosphorylated to activate its own kinase activity; (4) DNA-PKcs acts as an adhesive to join both broken ends of the DNA to prevent DNA degradation by exonuclease; (5) the DNA is processed to remove non-ligatible ends or other forms of damage at the breaks; and (6) the XRCC4-DNA ligase IV complex repairs the DNA ends (in some cases, DNA polymerase may be required to synthesize new ends prior to ligation). When DNA PKcs is phosphorylated, it can induce conformational changes of proteins and regulate the activities of various proteins (such as Artemis, Ku70, Ku80, the DNA ligase) during NHEJ, which is crucial to the DNA repair process. Therefore, phosphorylated DNA PKcs (pDNA-PKcs) is often used as a marker of cellular DSBs.
Studies have shown that DNA-PK activity is associated with occurrence and development of various tumors. For example, in melanoma, DNA-PKcs can promote angiogenesis and tumor metastasis; in multiple myeloma, DNA-PKcs expression is significantly up-regulated; and in radiotherapy-resistant thyroid tumors, the content of Ku protein is significantly increased (Ihara, Ashizawa, Shichijo, & Kudo, 2019). Therefore, the combination of a DNA-PK inhibitor with an anti-tumor therapy that causes DNA damage (such as IR, chemotherapeutic agents, etc.) may be contemplated to improve the efficacy. The use of DNA-PK inhibitors may interfere with the DNA repair function of normal cells to a certain extent. However, there are multiple complementary DNA repair pathways in normal cells, while tumor cells face strong DNA replication pressure and lack effective DNA repair means. Inhibition of the activity of DNA-PK in tumor cells can enhance the killing effect of other anti-tumor drugs against the tumor cells. For example, NU7441 can significantly enhance the efficacy of chemotherapy drugs that cause DNA damage against breast cancer and non small cell lung cancer, and can also improve the sensitivity of colorectal cancer to IR and chemotherapy.
Through years of research, a number of DNA-PK inhibitors have been discovered. The first compound discovered with DNA-PK kinase inhibitory activity was a fungal metabolite, Wortmannin, with an IC50 (DNA-PK) of about 15 nM, which also plays an important role in the acetylation and phosphorylation of the p53 protein (Sarkaria et al., 1998). A quercetin derivative LY294002 reported later also has DNA-PK inhibitory activity (Maira, Stauffer, Schnell, & Garcia-Echeverria, 2009). Afterwards a new generation of DNA-PK inhibitors such as NU7026 and NU7441 were developed based on the structure of LY294002. Although it has been confirmed that these compounds show a good killing effect on tumor cells, they have problems such as high toxicity and poor selectivity, and thus cannot enter clinical trials (Maira et al., 2009). Other DNA PK inhibitors have also been reported, such as small molecule compounds including
OK1035, SU11752, PP121, and KU-0060648, but these compounds also have disadvantages such as low specificity for DNA-PK (George et al., 2019). Therefore, there is still a need to develop DNA-PK inhibitors with high activity, high specificity, and low toxicity that better meet the clinical demands.
Summary of Invention
One or more embodiments of the present disclosure provide novel imidazolidinone derivatives, or stereoisomers, solvates, metabolites, pharmaceutically acceptable salts, co-crystals, or prodrugs thereof, a pharmaceutical composition comprising the same, and their use as a DNA-PK inhibitor.
The compounds according to one or more embodiments of the present application have high inhibitory activity against and high selectivity for DNA-PK, and can be used as a sensitizer for chemotherapy and radiotherapy to prevent and/or treat cancer, improve the therapeutic efficacy, and reduce toxic and side effects.
One or more embodiments according to the present disclosure provide a compound of general formula (I), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, Ro
R3 NN O
(R C NH N
(I) wherein R3R
(R2 C (R 2 (R 2 C3 Br(R2R is ,or
--- is a single bond or a double bond;
A, R3 R2
A, B, C, and D in the are each independently C or N, and at least one of A, B, C, and D is N; Ro is H, C 1-6 alkyl, or cyclopropyl, wherein the C1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
0O Ri is 0, 0 , 0, Rib , Rib
, m( ),+( Rl Rl 0O. N Rl1a , or pyridyl, and Ri is 1b Rib optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, C1-6 alkyl, and C1-6 alkoxy; Ria is H or C1-6 alkyl; Rib is H, OH, cyano, or hydroxyl-substituted C 1-6 alkyl; R 2 is H, cyano, =0, carboxyl, -C(=)NR2aR2b, C1-6 alkoxy, C1-6 alkyl, halogen, S(=0) 2R2 a or -C(=O)OCI-6 alkyl; wherein the C1-6 alkyl, -C(=)OC-6 alkyl or C 1-6 alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are each independently H, C 1-6 alkyl or 3- to 5-membered cycloalkyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from OH, D, halogen, C1-6 alkyl and C1-6 alkoxy; alternatively, R2 aand R2b together with the atom to which they are attached form 5- to 6-membered heterocyclyl group containing 1, 2 or 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from C1-6 alkyl, OH, and halogen; R3 is halogen or C1-6 alkyl, wherein the C1-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; x and y are each independently 1, 2, or 3; provided that R3
(R2 C Ri is not , , or OH when , Ro, R 2, and R3 all satisfy the following condition: R3 R3
(R 2 (RC is , n is 1, Ro is H or methyl, R2 is methoxy or S(=0)2Me, and R3 is methyl.
In one or more embodiments according to the present disclosure, R1 is OH,
F OH OH F OH CN,
D OH CN CN 0, 0
0 to N, 0I 4 ORia iOH O
, OH OH OH , OH , : 60
O N ,O, CN, or ; Ria is H or C1 -6alkyl; R2 is H, cyano, -C(=0)NR2aR2, C1 -6alkoxy, halogen, -S(=0) 2 R2a or -C(=O)OCi.6 alkyl; wherein the -C(=O)OC1-6 alkyl or C1 -6 alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are each independently H, C 1-6 alkyl or 3- to 5-membered cycloalkyl, wherein the CI-6 alkyl is optionally further substituted with one or more substituents selected from OH, D, and halogen; alternatively, R2a and R2b together with the atom to which they are attached form 5- to 6-membered heterocyclyl group containing 1to 3 heteroatoms selected from N, 0 and S, wherein the heterocyclyl group is optionally further substituted with one or more substituents selected from OH and halogen; R3 is halogen or C1-6 alkyl, wherein the Ci-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; provided that R3
Ri is not , , or OH when , Ro, R2, and R3 all satisfy the following condition:
R3
(R2R is(, n is 1, Ro is H or methyl, R2 is methoxy or S(=0)2Me, and R3 is methyl.
In one or more embodiments according to the present disclosure, R3
(R2is (R2 or 2)R
Ro is H, Ci-4 alkyl, or cyclopropyl, wherein the Ci-4 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
N 0 Ri is 0 0 R1a OH CN,
D O ,CN, O ',OH, Oor 0; Ria is H, C1-6 alkyl or -C(=O)C1-6 alkyl; R2 is H, cyano, -C(=0)NR2aR2, C1 -6 alkoxy, halogen, -S(=0) 2R2a or -C(=O)OCi.6 alkyl; wherein the -C(=O)O alkyl or C1 -6 alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2 a and R2b are each independently H, C 1-6 alkyl or 3- to 5-membered cycloalkyl, wherein the C 1.6 alkyl is optionally further substituted with one or more substituents selected from OH, D, and halogen; alternatively, R 2 aand R2b together with the atom to which they are attached form 5- to 6-membered heterocyclyl group containing 1to 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from OH and halogen; R3 is halogen or C1-6 alkyl, wherein the C1-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; provided that R3
(R C Ri is not , ,or OH when ,Ro, R 2, and R3 all satisfy the following condition:
R3
(R - C (R 2 rR is , n is 1, Ro is H or methyl, R2 is methoxy or S(=0)2Me, and R3 is methyl.
In one or more embodiments according to the present disclosure, the compound according to the present disclosure is a compound of general formula (II):
R2 d ~a RO R3 N`
R2C N N N H R1 (II) wherein Ro is H, C1 -6 alkyl, or cyclopropyl, wherein the Ci-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
Ri is 0, 0 , 0 Rib Rib
N R Ri Rb 1lb ib 1a or pyridyl, and R1 is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, CI-6 alkyl, and CI-6 alkoxy; Ria is H or C1 -6 alkyl; Rib is H, OH, cyano, or hydroxyl-substituted C1 -6 alkyl; R2c is H, cyano, halogen, or C 1-6 alkoxy; R2d is H, cyano, carboxyl, -C(=0)NR2aR2b, C1-6 alkyl, halogen, -S(=0) 2R2 a or C(=O)OC 1-6 alkyl; wherein the C1 -6 alkyl or -C(=O)OC1 -6 alkyl is optionally substituted with one or more substituents selected from halogen and deuterium; R2 a and R2b are H, C1-6 alkyl or 3- to 5-membered cycloalkyl, wherein the C1.6 alkyl is optionally further substituted with one or more substituents selected from OH, D, halogen, C 1-6 alkyl and CI-6 alkoxy; alternatively, R 2 aand R2b together with the atom to which they are attached form 5- to 6-membered heterocyclyl group containing 1to 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from CI.6 alkyl, OH, and halogen; R3 is halogen or C1-6 alkyl, wherein the C1-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; x and y are each independently 1, 2, or 3; provided that R3
Ri is not , ,or OH when ,Ro, R 2, and R3 all satisfy the following condition: R3
n (R C (R 2 R3
. is selected from , n is 1, Ro is H or methyl, R2 is methoxy or -S(=0)2Me, and R 3 is methyl.
In one or more embodiments according to the present disclosure, the compound according to the present disclosure is a compound of general formula (III): 0 RO R2 a\ N R 2 b-NN R 2C N N N H R1 (J) wherein Ro is H, C1 -6 alkyl, or cyclopropyl, wherein the Ci-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
0O Ri is 0, 0 , 0, Rib Rib
Rib Rib (~O)( C-N, o n~i Rlb Rlb ' Ria or pyridyl, and R1 is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, CI-6 alkyl, and CI-6 alkoxy; Rib is H, OH, cyano, or hydroxyl-substituted C1 -6 alkyl; R2 a and R2b are each independently H, C 1-6 alkyl or 3- to 5-membered cycloalkyl, wherein the CI-6 alkyl is optionally further substituted with one or more substituents selected from D and halogen; alternatively, R2 aand R2b together with the atom to which they are attached form 5- to 6-membered heterocyclyl group containing 1to 3 heteroatoms selected from N, 0 and
S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from C 1-6 alkyl, OH, and halogen; R2, is H, cyano, halogen, or C 1-6 alkoxy, wherein the C 1-6 alkoxy is optionally substituted with one or more D; R3 is halogen or C 1-6alkyl, wherein the C 1-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; x and y are each independently 1, 2, or 3.
One or more embodiments according to the present disclosure provide a compound of general formula (IV), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, 0 Ro H 2N N N F N N H R1 (IV) wherein Ro is H, C 1-6 alkyl, or cyclopropyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium; and R 1 is a -(CH)m- 4 to 7-membered carbocyclyl, -(CH)m- 4 to 7-membered heterocyclyl, -(CH)m- 8 to 12-membered bridged ring, or -(CH)m- 7 to 12 membered spiro, wherein the -(CH)m- 4 to 7-membered carbocyclyl, -(CH)m- 4 to 7-membered heterocyclyl, (CH)m- 8 to 12-membered bridged ring, or -(CH)m- 7 to 12 membered spiro is optionally further substituted with one or more substituents selected from hydroxy, cyano, halogen, =0, C 1 -6 alkyl, C 1-6 alkoxy, and hydroxyl-substituted C 1 -6alkyl.
In one or more embodiments according to the present disclosure, Ro is H, C1 -6 alkyl, or cyclopropyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
Ri is a, , , , , Rib Rib
Ri Ri NoR ad -i 1lb 1a ib or pyridyl, and R1 is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, C 1-6alkyl, and C 1-6alkoxy;
Ria is H or C 1 .6 alkyl; Rib is H, OH, cyano, or hydroxyl-substituted C 1 .6 alkyl; m is 0 or 1; and x and y are each independently 1, 2, or 3.
In one or more embodiments according to the present disclosure, Ro is C1 .4 alkyl, wherein the C 1 .4 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium; and
Ri is CN OH OH OH
F CN CN F OH OH F
0, )>o NH, OH 0
OH CN O 0 OH O
OH, OHor N
In one or more embodiments according to the present disclosure, the compound according to the present disclosure is F3 C O /F O CI O O/O N>=N )= all N N NN N N N N ON N N N H 'N N H H H H 00 0 0
HN N N H 2N O
NN N N< N N NA NN F NNN N H H H 0 0, 0 H 0 0 N 0/ N ~ ~N NHN ' CIfN/ N N N FN N N N" N Ho H F H
00 0
C1 /, 0 F /N~ F N' N 2 ' 'N"' N' N>=
N N N NN N N N N F H bH bH 0 0 0
H2 N CN N Nl N' N' N"' : ~~ 1 N> N~N N N0: a N '1 _ H bH bHb 0 0 0 0 /N~ 0/
H2N N " N" "'N" )!~ >zzON- L )== H 0L>ZZ "
" N N N N N N N N H bH Hb
0 0 0 0 /0 /0/ N ' N 'N'N N'N -N "
" H" H Z 0L H' NIN -N>= NI N N N N N H bH bH 0 0 0
N N 3N N'N 'HA >zz0H 2N N'NA N N/NI >=o N N N H 'o'- N N N H b ~Hb 00 0 N N~ N N
H :N>==0N
' N'N =H N" N>=
NH Hb
0, N OH 0,
NN N CN NN N/ ~N~n' N. N' NNN 'N N NN AL> >= H H N~ H 00 0 0 0 0 H2N N' -- N N 0 / '
F N "N N 2 N N N 0 /
N NL N>=NN = H N ~F N HN N -N HH
N 00 0/
H2N N' NN' N0
F Nq N N NN'N 0 N' N' HH `jN N:> F N N N/ N N N N H bHb N 0 0
0 0 D3C,. N >= N N N N NNN N 0 N N N NN H 11K- N 1O iio 2NiX N" NN =o H b H bH bHb 0 0 0 0
H N 0 ZZ N~I N N~~N NN NN ,-NN N Nj N>=
0 0 0 0
I >o II== N N H N 0N ~N H H
00 0 0
O0 S- jlNN = S fN NN N N H 1 N =0H - A Nn) = 0 NN N N0 N N N N N 2 H boH bHb O0 0
H2 N N _ > 2 N~ S >0 'N > ~~ N : -1N N "-NNN, N NINN NIN N N N N H boH bHb O0 0
H0-2NNN N>= HN-' N- HO == -N N NN=
N H )~ ZZ ) >= N N NN N NN N N> H2 - N H H N H b 0 0 0 0 0 0
H 2N -N H2N -~N N ' 0 FC~" N N ON , FNN N H H H b 0 0 0 0 0 00 HON I N: llN>= H2 - N N > 1 0 N N) H2H`' N-OH N--' >=:---:>= FC N 'N N A 0 N N NNN N N NN N H H H Hb
O0 0 0 0 0 0 CIN H2N ,
N N N. N!'N)INF= N N N F N N, N H H H b 0O 0 0 0 0 0
H 2N I~ N 2N N N H2Nt]N NN N F- H N ; N F N N >-N F AN N N H bH bHb 0 0 N
0 0 \r12
0 0 0 H2 NH2 N'I NN ~F NN N= H 2N' N NN N= - )N> IZZ j F N 'NN H O bH OH H N N F N N N
b
H2OH
F j.,f =oH 2N N ,-NN N H2Nt:-_ N N N ~ H N F N NN F N N N
H H H b
0- OH H 0 0 0 H 2N N N
H 2N N NN N F N N N >= H 2N n N N 0 H F N N NF N N N
F -OH
00 /H 2N NHN N N== 2 NN H2N N F N- N \'N F NO HOH
O OH 0 0 0 ~ H2N N - ~~>=o NN N H2N j N-- 4PN >N H2Nt - - N )9>=0N N H F N N N F
OH 0ON 0 0 0/ NN N NHN N>=N
F NN N I .F N NN H FC N N N H
O -H HOb
00/
F N~ N HN N N
H NNN H H N ,N 0
0oO H 0 OH
0F 0 D
H 2N N N H 2Nt N N H2Nt N NN F N N N F N N N F ~N N N H H H F-b
bo 0 0
F 0 0 f OrFH 2Nt N N-: N N= N2N N==
H2N N N N 0 F N N N F N 'N N
N N NHH
0o N N
F 0 0 F-j 0 H 2 N~O N H2N O _0N~ OzzO 'aCN N >z N N N DN N HNN H O H
kH, ~ 0 or b
One or more embodiments according to the present disclosure provide a pharmaceutical composition comprising: (1) a compound according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, (2) optionally one or more additional active ingredients; and (3) a pharmaceutically acceptable carrier and/or excipient.
One or more embodiments according to the present disclosure provide use of the compound according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the pharmaceutical composition according to the present disclosure, in the manufacture of a medicament for treatment of cancer.
In one or more embodiments according to the present disclosure, the medicament for treatment of cancer is a DNA-PK inhibitor.
One or more embodiments according to the present disclosure provide a compound of general formula (I'), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, Ro
R3 NN O
(Rn C NH (I') wherein C is selected from phenyl and 6-membered monocyclic heterocyclic group, wherein the heterocyclic group may contain 1 to 3 heteroatoms selected from N; Ro is selected from H,C1 .6 alkyl, and cyclopropyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
mo ,
R1 is selected from O, 0 0, , Ria ,
D OH CN CN CN O' OH
O 0 and 0 Ria is selected from H, C1-6 alkyl, and -C(=O)C1-6 alkyl; R2 is selected from cyano, carboxyl, -NR2aR2b, -C(=0)NR2aR2b, C1-6 alkoxy, C1-6 alkyl, halogen, -S(=0)2 R2 a and -C(=O)OC 1 -6 alkyl; wherein the alkyl or alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C1-6 alkyl and 3- to 5-membered cycloalkyl, wherein the C1 -6 alkyl is optionally further substituted with one or more substituents selected from OH, D, halogen, C1 -6 alkyl and CI-6 alkoxy; R2a and R2b together with the atom to which they are attached may form 5- to 6 membered heterocyclyl group which may contain 1 to 3 heteroatoms selected from N, O and S, wherein the heterocyclyl group may be further substituted with one or more substituents selected from C 1-6alkyl, OH, and halogen; R3 is selected from halogen and C1 -6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; is selected from 0 and 1; and n is selected from 0, 1, and 2; provided that m
R 1 is not 0 or OH when Cis selected from phenyl and a 6-membered monocyclic heterocyclic group and Ro is H.
In one or more embodiments according to the present disclosure, R3
(R2 Ro (R2 R is or Ro is selected from H, C 1-4alkyl, and cyclopropyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
Ri is selected from R , O 0, , Ra,
D OH CN CN, CN, O' OH
O and 0 Ria is selected from H,C1 .6 alkyl, and-C(=O)C 1 -6alkyl; R2 is selected from cyano, -C(=0)NR2aR2, C 1.6 alkoxy, halogen, -S(=0)2R2a and 1 alkyl; wherein the alkyl or alkoxy is optionally substituted with one or C(=)OC -6 more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C1 .6 alkyl and 3- to 5-membered cycloalkyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from OH, D, and halogen; R2 a and R2b together with the atom to which they are attached may form 5- to 6 membered heterocyclyl group which may contain 1 to 3 heteroatoms selected from N, O and S, wherein the heterocyclyl group may be further substituted with one or more substituents selected from OH and halogen; R3 is halogen or C1 -6alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is selected from 0 and 1; and n is selected from 0, 1, and 2; provided that m
R 1 is not 0 or OH when Cis selected from phenyl and a 6-membered monocyclic heterocyclic group and Ro is H.
One or more embodiments according to the present disclosure provide a compound of general formula (I"), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, Ro
(R R NH N O (I") wherein C is selected from phenyl and 6-membered monocyclic heterocyclic group, wherein the heterocyclic group may contain 1 to 3 heteroatoms selected from N; Ro is selected from H and C1 .6 alkyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
R 1 is selected from O, O, 0, OH , CN , and
0--' R2 is selected from cyano, carboxyl, -NR2aR2b, -C(=0)NR2aR2b, C 1-6 alkoxy, C1-6 alkyl, halogen, -S(=0) 2 R2 aand -C(=)OC 1-6 alkyl; wherein the alkyl is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C 1-6alkyl or 3- to 5-membered cycloalkyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from OH, halogen, C 1 -6alkyl and C 1-6alkoxy; R2 a and R2b together with the atom to which they are attached may form 5- to 6 membered heterocyclyl group which may contain 1 to 3 heteroatoms selected from N, O and S, wherein the heterocyclyl group may be further substituted with one or more substituents selected from C 1-6 alkyl, and halogen; R3 is selected from halogen and CI.6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is selected from 0 and 1; and n is selected from 1 and 2.
In one or more embodiments according to the present disclosure, R3
(R 2 c (R2 R3 (R2) R is selected from or Ro is selected from H and C1 -4 alkyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
R1 is selected from R1 is selected from 0 0 ,
OH ,CN and R 2 is selected from cyano, -C(0)NR2aR2b, CI-6 alkoxy, halogen, -S(=0)2R2a and C(=)OCi-6 alkyl; wherein the alkyl is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C1-6 alkyl and 3- to 5-membered cycloalkyl, wherein the C1 -6 alkyl is optionally further substituted with one or more substituents selected from OH and halogen; R2 a and R2b together with the atom to which they are attached may form 6-membered heterocyclyl group; R3 is selected from halogen and C1 -6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is selected from 0and 1; and n is selected from Iand 2. In one or more embodiments according to the present disclosure, the compound according to the present disclosure is
NNAN N NAN
b H bH 0 0 0 00
0== 0SC>= - N- NH N N HNNN N
H N~ N=0 N>SZO ~NN H H
0 0 0 0 0 NC N N H2 N - . N ~ N N N> >==o >-(: 'jN NN b H bHb 0 0 0
N \-o H2 N ~CI Nll N>== ~ C N N ~ NN N N NN N F N N NN H bF H bF Hb
0 0 0 0 F / ~F 0 c
H 2N N N'- N NH 2N N N NA N N NN nN/ NNANN H bH bHb 0 0 0 0 0
N ~ N N NCaC NINN N 2
~N N N H bH bHb 0 0 0 N 0 /0/ ~ ~N N - NN- N N~ >=0 HN : N >= --- N- * - N- N \= N NN H bH bHb 0 0 0, 0 0/ AN N '- N.N N * NNN 0/ A' I >0== H 2N '
N HH
0 ,0
H 2 N /NC ~ "- "- Y/-D ~. N:l CD N- N HNNa--_ N>= N >= N N N N N
NC) nN N" N/ H N N
NCO H> Ca N" N>= N 1 >= N N q-N H'b NNNNH NC / N
H HH N N N N N N HH
N N N H >=
CN 11N H2 \ 0 H 2N N N- N H 2NN' t" N NN F N N F\AN"N F N N N H
bo CN, CON,
0 0 H 0 ' " 0 / NO 0 F - N'" N11 _ HN I \- N H N N F A N '"'N N N HHO
0N 0 , 0 00 N' N- 2N ' N' N H >= - N>=OHN N N N F AN" NNF N N N N H H NH"
0 / 0 0
D3C N H2N- N 'NN' 0 "
1, >= N NF' NN N N-NN'N'N N N N H H H NHN >=
0o 0 N "N N N' 0" NN NN "N"" - N N N~ N rN NI NN" N N NN N NN N N N
H 2 0- N NN'- N N= H' N/anN= N N N N
0r 0N
N' 3C N' 19
One or more embodiments according to the present disclosure provide a pharmaceutical composition comprising: (1) a compound according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, (2) optionally one or more additional active ingredients; and (3) a pharmaceutically acceptable carrier and/or excipient.
One or more embodiments according to the present disclosure provide use of the compound according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the pharmaceutical composition according to the present disclosure, in the manufacture of a DNA-PK inhibitor.
One or more embodiments according to the present disclosure provide the compound of general formula (I), (II), (III), (IV), (I'), or (I") or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the composition according to the present disclosure, for use as a medicament.
One or more embodiments according to the present disclosure provide the compound of general formula (I), (II), (III), (IV), (I'), or (I") or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the composition according to the present disclosure, for use as a medicament for treatment of cancer or for use in a method for preventing and/or treating cancer.
One or more embodiments according to the present disclosure provide the compound of general formula (I), (II), (III), (IV), (I'), or (I") or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the composition according to the present disclosure, for use as a DNA-PK inhibitor or for use in a method for inhibiting DNA-PK.
One or more embodiments according to the present disclosure provide the compound of general formula (I), (II), (III), (IV), (I'), or (I")or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the composition according to the present disclosure, for use in treatment of DNA-PK-associated diseases.
One or more embodiments according to the present disclosure provide a method for inhibiting DNA-PK, comprising contacting the compound of general formula (I), (II), (III), (IV), (I'), or (I") or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co crystal, or prodrug thereof, or the composition according to the present disclosure, with a subject in need thereof.
One or more embodiments according to the present disclosure provide a method for treating DNA-PK-associated diseases, comprising administering the compound of general formula (I), (II), (III), (IV), (I'), or (I") or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the composition according to the present disclosure, to a subject in need thereof.
One or more embodiments according to the present disclosure provide a method for treating cancer, comprising administering the compound of general formula (I), (II), (III), (IV), (I'), or (I") or the above specific structures according to the present disclosure, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co crystal, or prodrug thereof, or the composition according to the present disclosure, to a subject in need thereof. Unless stated to the contrary, the terms used in the specification and claims have the following meanings. Carbon, hydrogen, oxygen, sulfur, nitrogen, F, Cl, Br and I involved in the groups and compounds described herein are each inclusive of isotopes thereof, and carbon, hydrogen, oxygen, sulfur or nitrogen involved in the groups and compounds described herein is optionally further replaced by one or more isotopes thereof corresponding thereto, wherein isotopes of carbon include 1 2 C, "C and 14 C, isotopes of hydrogen include protium (H), deuterium (D, also called heavy hydrogen) and tritium (T, also called superheavy hydrogen), isotopes of oxygen include 160, 170 and 180, isotopes of sulfur include 3 2 s, 3 3 , 3 4 S and 3 6S, isotopes of nitrogen include 14N and "N, isotopes of fluorine include 1 7 F and 19F, isotopes of chlorine include3 C1 and 37 C1, and isotopes of bromine include 79Br and8 1 Br. "Alkyl" refers to a linear or branched saturated aliphatic hydrocarbon group consisting of I to 20 carbon atoms, preferably an alkyl group consisting of I to 8 (e.g., 1, 2, 3, 4, , 6, 7 or 8) carbon atoms, more preferably an alkyl group consisting of 1 to 6 carbon atoms, and further preferably an alkyl group consisting of 1 to 4 carbon atoms. Non limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl and various branched chain isomers thereof; when the alkyl is substituted, it may be optionally further substituted with 1 or more substituents. "Alkoxy" refers to a group formed by substitution of at least 1 carbon atom of an alkyl group with an oxygen atom. Non-limiting examples include methoxy, ethoxy, n propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropoxy and cyclobutoxy. The alkyl is defined in the same way as for the "alkyl" described above. "Alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group containing 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon-carbon double bonds and consisting of 2 to 20 carbon atoms, preferably an alkenyl group consisting of 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12) carbon atoms, more preferably an alkenyl group consisting of 2 to 8 carbon atoms, and further preferably an alkenyl group consisting of 2 to 6 carbon atoms. Non-limiting examples include vinyl, propen-2-yl, buten-2-yl, penten-2-yl, penten-4-yl, hexen-2-yl, hexen-3-yl, hepten-2-yl, hepten-3-yl, hepten-4-yl, octen-3-yl, nonen-3-yl, decen-4-yl, and undecen-3-yl. The alkenyl may be optionally further substituted with 1 or more substituents. "Alkynyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group containing 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon-carbon triple bonds and consisting of 2 to 20 carbon atoms, preferably an alkynyl group consisting of 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12) carbon atoms, more preferably an alkynyl group consisting of 2 to 8 carbon atoms, and further preferably an alkynyl group consisting of 2 to 6 carbon atoms. Non-limiting examples include ethynyl, propyn-1-yl, propyn-2-yl, butyn-1-yl, butyn-2-yl, butyn-3-yl, 3,3-dimethylbutyn-2-yl, pentyn-1-yl, pentyn-2-yl, hexyn-1-yl, 1-heptyn-1-yl, heptyn-3-yl, heptyn-4-yl, octyn-3-yl, nonyn-3-yl, decyn-4 yl, undec-3-yl, and dodecyn-4-yl. The alkynyl may be optionally further substituted with one or more substituents. "Aryl" refers to a substituted or unsubstituted aromatic ring. It may be a 5-8 membered (e.g., 5, 6, 7 or 8 membered) monocyclic ring system, a 5-12 membered (e.g., 5, 6, 7, 8, 9, 10, 11 or 12 membered) bicyclic ring system or a 10-15 membered (e.g., 10, 11, 12, 13, 14 or 15 membered) tricyclic ring system, and may be a bridged ring or a spiro ring. Non-limiting examples include phenyl and naphthyl. The aryl may be optionally further substituted with 1 or more substituents. "Heteroaryl" refers to a substituted or unsubstituted aromatic ring. It may be a 3-8 membered (e.g., 3, 4, 5, 6, 7 or 8 membered) monocyclic ring system, a 5-12 membered (e.g., 5, 6, 7, 8, 9, 10, 11 or 12 membered) bicyclic ring system or a 10-15 membered (e.g., 10, 11, 12, 13, 14 or 15 membered) tricyclic ring system, and it contains I to 6 (e.g., 1, 2, 3, 4, 5 or 6) heteroatoms selected from N, 0 and S, and is preferably 5-8 membered heteroaryl. Ito 4 (e.g., 1, 2, 3 or 4) N and S optionally substituted in the ring of the heteroaryl can be oxidized to various oxidation states. Heteroaryl may be attached to a heteroatom or carbon atom and it may be a bridged ring or a spiro ring. Non-limiting examples include cyclic pyridinyl, furanyl, thienyl, pyranyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl, benzimidazolyl, benzopyridinyl and pyrrolopyridinyl. Heteroaryl is optionally further substituted with 1 or more substituents. "Carbocyclyl" or "carbocycle" refers to a saturated or unsaturated, aromatic or non aromatic ring. When being an aromatic ring, it is defined in the same way as for the "aryl" described above; when being an non-aromatic ring, it may be a 3-10 membered (e.g., 3,4,5,6, 7,8, 9 or 10 membered) monocyclic ring system, a 4-12 membered (e.g., 4, 5, 6, 7, 8, 9, 10, 11 or 12-membered) bicyclic ring system or a 10-15 membered (e.g., , 11, 12, 13, 14 or 15 membered) tricyclic ring system, and it may be a bridged ring or a spiro ring. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, 1 cyclopentyl-1-alkenyl, 1-cyclopentyl-2-alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, , and . The "carbocyclyl" or "carbocycle" is optionally further substituted with 1 or more substituents. "Heterocyclyl" or "heterocycle" refers to a saturated or unsaturated, aromatic or non aromatic heterocycle. When being an aromatic heterocycle, it is defined in the same way as for the "heteroaryl" described above; when being a non-aromatic heterocycle, it may be a 3-10 membered (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 membered) monocyclic ring system, a 4-12 membered (e.g., 4, 5, 6, 7, 8, 9, 10, 11 or 12 membered) bicyclic ring system or a 10-15 membered (e.g., 10, 11, 12, 13, 14 or 15 membered) tricyclic ring system, and it contains 1 to 4 (e.g., 1, 2, 3 or 4) heteroatoms selected from N, 0 and S, and is preferably 3-8 membered heterocyclyl. 1to 4 (e.g., 1, 2, 3 or 4) N and S optionally substituted in the ring of the "heterocyclyl" or "heterocycle" can be oxidized to various oxidation states; "heterocyclyl" or "heterocycle" may be attached to a heteroatom or a carbon atom, and may be a bridged ring or a spiro ring. Non-limiting examples of "heterocyclyl" or "heterocycle" include epoxyethyl, epoxypropyl, aziridinyl, oxetanyl, azetidinyl, thietanyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxanyl, azepanyl, oxepanyl, thiepanyl, oxoazepinyl, diazepinyl, thiazepinyl, pyridinyl, piperidinyl, homopiperidinyl, furanyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, oxathianyl, 1,3-dithianyl, dihydrofuranyl, dithiacyclopentyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridinyl, pyrrolopyridinyl, benzodihydrofuranyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 3H-indolylquinolizinyl, N-pyridylurea, 1,1 dioxothiomorpholinyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl, aza-adamantyl and oxaspiro[3.3]heptyl. The "heterocyclyl" or "heterocycle" may be optionally further substituted with 1 or more substituents. "Cycloalkyl" refers to a saturated cyclic hydrocarbon group, the ring of which may be a 3-10 membered (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 membered) monocyclic ring system, a 4-12 membered (e.g., 4, 5, 6, 7, 8, 9, 10, 11 or 12 membered) bicyclic ring system or a -20 membered (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 membered) polycyclic ring system. The ring carbon atoms are preferably 3 to 10 carbon atoms, further preferably 3 to 8 carbon atoms. Non-limiting examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, 1,5 cyclooctadienyl, 1,4-cyclohexadienyl, cycloheptatrienyl, and the like. When the cycloalkyl is substituted, it may be optionally further substituted with 1 or more substituents. "Heterocycloalkyl" refers to a substituted or unsubstituted saturated non-aromatic cyclic group. It may be a 3-8 membered (e.g., 3, 4, 5, 6, 7 or 8 membered) monocyclic ring system, a 4-12 membered (e.g., 4, 5, 6, 7, 8, 9, 10, 11 or 12 membered) bicyclic ring system or a 10-15 membered (e.g., 10, 11, 12, 13, 14 or 15 membered) tricyclic ring system, and it contains 1, 2 or 3 heteroatoms selected from N, 0 and S, and is preferably 3-8 membered heterocyclyl. 1, 2 or 3 N and S optionally substituted in the ring of "heterocycloalkyl" can be oxidized to various oxidation states; "heterocycloalkyl" may be attached to a heteroatom or a carbon atom and may be a bridged ring or a spiro ring. Non-limiting examples of "heterocycloalkyl" include epoxyethyl, aziridinyl, oxetanyl, azetidinyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3 dioxanyl, azepanyl, piperidinyl, morpholinyl, thiomorpholinyl, 1,3-dithianyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, azabicyclo[3.2.1]octyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecyl,aza-adamantylandoxaspiro[3.3]heptyl. When the "alkyl", "alkoxy", "alkenyl", "alkynyl", "aryl", "heteroaryl", "carbocyclyl", "carbocycle", "heterocyclyl", "heterocycle", "cycloalkyl", "heterocycloalkyl" or "heterocyclyl" described above is substituted, it may be optionally further substituted with 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 substituents selected from F, Cl, Br,I, hydroxy, mercapto, nitro, cyano, amino, C 1 .6 alkylamino,=0, C 1 .6 alkyl, C 1 .6 alkoxy, C2-6 alkenyl, C2-6 alkynyl, -NR =NR6, -C(=0)-OC kOC(=0)C-6 alkyl, -C(=)NR4 (4R5, 4 R5
, C3-8 cycloalkyl, C3.8 heterocycloalkyl, C6-10 aryl, C 5-10 heteroaryl, -C(=O)OC6-10 aryl, OC(=O)C6-io aryl, -OC(=O)C5-1o heteroaryl, -C(=O)OC 5-1 o heteroaryl, -OC(=O)C 3-8 heterocycloalkyl, -C(=O)OC 3 .8 heterocycloalkyl, -OC(=)C 3 - 8 cycloalkyl, -C(=0)OC 3
. 8 cycloalkyl, -NHC(=0)C 3 .8 heterocycloalkyl, -NHC(=0)C6-io aryl, -NHC(=0)C- 1 o heteroaryl, -NHC(=0)C 3 -8 cycloalkyl, -NHC(=0)C 3 .8 heterocycloalkyl, -NHC(=0)C 2 6 alkenyl and -NHC(=0)C 2 -6 alkynyl, wherein the substituent C1 -6 alkyl, C1 -6 alkoxy, C2-6 alkenyl, C2-6 alkynyl, C3-8 cycloalkyl, C38 heterocycloalkyl, C-10 aryl, C 5-10 heteroaryl, -NHC(=0)C6-io aryl, -NHC(=0)C5 -io heteroaryl, -NHC(=0)C 3 -8 heterocycloalkyl or -NHC(=0)C 3 -8 cycloalkyl is optionally further substituted with 1 to 3 substituents selected from OH, F, Cl, Br, I, C1 .6 alkyl, C1 .6 alkoxy, -NR4Rq5 and =0, Rq1 is selected from C 1 6 alkyl, C 16 alkoxy and C6.10 aryl, and Rg2 and Rq3 are selected from H and C 1 .6 alkyl, wherein Rq4 and Rq5 are selected from H, C 1.6 alkyl, NH(C=NR9l)NRq 2 Rq 3, -S(=0) 2NRg 2 Rq 3 , -C(=)R 1 and -C(=O)NRq 2Rq 3 , wherein the C 1-6 alkyl is optionally further substituted with 1 or more substituents selected from OH, F, Cl, Br, I, C1 .6 alkyl, C1 .6 alkoxy, C6.1o aryl, C 5-10 heteroaryl, C3-8 cycloalkyl and C3-8 heterocycloalkyl; or Rq4 and Rq5, together with an N atom, form a 3-8 membered heterocycle, which may contain 1 or more heteroatoms selected from N, 0 and S. "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof' refers to a salt obtained by reaction of a compound disclosed herein in a free acid form with a nontoxic inorganic or organic base or by reaction of a compound disclosed herein in a free base form with a nontoxic inorganic or organic acid, and in this salt, the bioavailability and characteristics of the compound disclosed herein in the free acid or free base form is retained. "Pharmaceutical composition" refers to a mixture of one or more compounds described herein or pharmaceutically acceptable salts or prodrugs thereof and other chemical components, wherein the "other chemical components" refer to pharmaceutically acceptable carriers, excipients and/or one or more other therapeutic agents.
"Carrier" refers to a material that does not cause significant irritation to an organism and does not eliminate the biological activity and characteristics of the administered compound. "Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders and disintegrants. "Prodrug" refers to a compound disclosed herein that can be metabolized in vivo to become biologically active. A prodrug disclosed herein is prepared by modifying amino or carboxyl in a compound disclosed herein, and the modification can be removed by conventional operation or be removed in vivo to obtain the parent compound. When a prodrug disclosed herein is administered to a mammalian subject, the prodrug is cleaved to form free amino or carboxyl. "Cocrystal" refers to a crystal formed by binding of an active pharmaceutical ingredient (API) and a cocrystal former (CCF) via a hydrogen bond or other non-covalent bonds, wherein the API and CCF are both solid in their pure state at room temperature, and the components are present in a fixed stoichiometric ratio. A cocrystal is a multi-component crystal, including both a binary cocrystal formed by two neutral solids and a multiple cocrystal formed by a neutral solid and a salt or solvate. "Stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and conformers. "Optional", "optionally", "selective" or "selectively" means that the subsequently described event or circumstance may, but does not necessarily, occur, and the description includes cases where the event or circumstance occurs and cases where it does not. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl may, but does not necessarily, be present, and the description includes the case where the heterocyclyl is substituted with alkyl and the case where the heterocyclyl is not substituted with alkyl. DETAILED DESCRIPTION The following examples illustrate the technical schemes of the present invention in detail, but the protection scope of the present invention includes but is not limited thereto. DMSO: dimethyl sulfoxide; DTT: dithiothreitol; ATP: adenosine triphosphate; DNA: deoxyribonucleic acid; IC50: refers to the concentration of a compound at which the activity of DNA-PK kinase is 50% inhibited; MPLC: medium-pressure silica gel column chromatography. The reagents used in the present application are all commercially available. Examples Intermediate 1 2-Chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (Intermediate 1)
N' - N Cl O CIN N
0 Intermediate 1
O 0
Step 1 CI O Step 2 N N OH 0 ` N CI N NH CI C7N)C N CI
O0 1A 1B 1C
H N
Step 3 CI -=O Step 4 NNN NC CI N
0 0 ID Intermediate 1
Step 1: Ethyl 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate (1B)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (30.00 g, 136.4 mmol) and tetrahydro 2H-pyran-4-amine hydrochloride (18.66 g, 136.4 mmol) were dissolved in acetonitrile (600 mL), and after the solution was stirred several times, potassium carbonate (46.92 g, 340.9 mmol) was added. The reaction mixture was stirred at room temperature for 4 h and monitored by TLC. After the reaction is complete, The mixture was filtered, and the filter residue was washed with ethyl acetate (300 mL). The filtrate was concentrated to give a crude product, which was then purified by column separation (n-hexane:ethyl acetate (v/v) = 1:1) to obtain the title compound ethyl 2-chloro-4-((tetrahydro-2H pyran-4-yl)amino)pyrimidine-5-carboxylate 1B (white solid, 30.0 g, 77.4% yield). 1H NMR (400 MHz DMSO) 6 8.62 (s,1H), 8.32 (d, 1H), 4.30 (q, 2H), 4.21-4.16 (in, 1H), 3.86-3.83 (in, 2H), 3.48-3.42 (in, 2H), 1.88-1.85 (m, 2H), 1.62-1.53 (in, 2H), 1.31 (t, 3H). LC-MS m/z(ESI) = 286.10 [M+1] Step 2: 2-Chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid (1C)
Ethyl 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate 1B (30 g, 104.99 mmol) was dissolved in tetrahydrofuran/water (200 mL/200 mL), and lithium hydroxide (5.03 g, 209.99 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 6 N hydrochloric acid, and a white solid was precipitated. The reaction mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to obtain the title compound 2-chloro-4-((tetrahydro-2H-pyran-4 yl)amino)pyrimidine-5-carboxylic acid 1C (white solid, 15.0 g, 55.44% yield).
'H NMR (400 MHz DMSO) 6 8.60 (s, 1H), 8.54 (d, 1H), 4.20-4.15 (m, 1H), 3.86-3.83 (m, 2H), 3.48-3.42 (m, 2H), 1.89-1.86 (m, 2H), 1.60-1.50 (m, 2H). LC-MS m/z(ESI) = 258.10 [M+1] Step 3: 2-Chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (1D)
2-Chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid IC (15 g, 58.21 mmol) was dissolved in dimethylacetamide (150 mL), and triethylamine (7.38 mL, 58.21 mmol) and diphenylphosphoryl azide (12.06 mL, 58.21 mmol) were added. The reaction mixture was then gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, and dried and concentrated in vacuo to give the title compound 2-chloro-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1D (white solid, 13.0 g, 87.69% yield). 1H NMR (400 MHz DMSO) 6 11.63 (s, 1H), 8.11 (s, 1H), 4.43-4.37 (m, 1H), 3.98-3.94 (m, 2H), 2.59-2.38 (m, 2H), 1.73-1.65 (m, 2H). LC-MS m/z(ESI)= 255.10 [M+1] Step 4: 2-Chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (intermediate 1)
2-Chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one D (5 g, 19.63 mmol) was dissolved in dimethylformamide (50 mL), dimethyl sulfate (2.48 g, 19.63 mmol) and cesium carbonate (9.5 g, 29.445 mmol) were added at 0 °C, and the reaction mixture was stirred at 0 °C for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The reaction mixture was filtered and collected to obtain the title compound 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 one intermediate 1 (white solid, 3.0 g, 56.87% yield). 1H NMR (400 MHz DMSO) 6 8.36 (s, 1H), 4.50-4.41 (m, 1H), 3.99-3.95 (m, 2H), 3.48
3.42 (m, 2H), 3.34 (s, 3H), 2.47-2.38 (m, 2H), 1.70-1.66 (m, 2H). LC-MS m/z(ESI) = 268.10 [M+1]
Intermediate 2 4-Amino-2-fluoro-5-methylbenzamide (intermediate 2)
0
H 2N
F NH 2
Intermediate 2
0 NC]I, stepi1
F NH 2 H2N NIH 2
2A Intermediate 2
4-Amino-2-fluoro-5-methylbenzonitrile 2A (300 mg, 2 mmol) and potassium carbonate (41.4 mg, 0.3 mmol) were dissolved in 1 mL of dimethyl sulfoxide, and 300 pL of hydrogen peroxide was added in an ice bath. The reaction mixture was gradually heated to 60 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, 5 mL of water was added to the reaction mixture, and a white solid was precipitated. The reaction mixture was filtered and concentrated under reduced pressure to yield the title compound 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (white solid, 160 mg, 47% yield). 'H NMR (400 MHz DMSO) 6 7.37 (d, 1H), 7.15 (s, 1H), 6.96 (s, 1H), 6.32 (d, 1H), 5.70 (s, 1H), 2.01 (s, 3H). LC-MS m/z(ESI)= 169.10 [M+1]
Example 1 7-Methyl-2-((2-methyl-4-(methylsulfonyl)phenyl)amino)-9-(tetrahydro-2H-pyran-4 yl)-7,9-dihydro-8H-purin-8-one (compound 1)
0 S N N N N >= H
0 Compound 1 I 0I 0 N N N S ,I X=0 -a >=
HNH2 00 Ia Intermediate 1 Compound 1 Compound la (290 mg, 1.08 mmol), compound intermediate 1 (200 mg, 1.08 mmol), cesium carbonate (703 mg, 2.16 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (98 mg, 0.11 mmol) were dissolved in dioxane (10 mL), followed by nitrogen purging. The resulting mixture was stirred under nitrogen at 100 °C for 4 h and monitored by TLC. After completion of the reaction, the mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to give the title compound 7-methyl-2-((2-methyl-4-(methylsulfonyl)phenyl)amino)-9-(tetrahydro 2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 1 (white solid, 50 mg, 11.09 %
yield). 'H NMR (400 MHz DMSO) 6 8.67 (s, 1H), 8.15 (s, 1H), 8.11 (d, 1H), 7.20 (d, 1H), 7.67 (dd, 1H), 4.47-4.39 (m, 1H), 3.98 (dd, 2H), 3.42 (t, 2H), 3.32 (s, 3H), 3.16 (s, 3H), 2.57-2.49 (m, 2H), 2.37 (s, 3H), 1.67 (dd, 2H). LC-MS m/z(ESI)= 418.10 [M+1]
Example 2 3-Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide (compound 2)
H2 N N N N _' . NN N>= H
Compound 2 I 0I 0 N" N H2 N ' N"'- N H2N + CI N N N N N
NH2 0 2a Intermediate 1 Compound 2
The title compound 3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide compound 2 (white solid, 100 mg, 19.64% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.48 (s,1H), 8.10 (s,1H), 7.82 (d, 2H), 7.71 (s, 1H), 7.66
(dd, 1H), 7.16 (s, 1H), 4.44-4.37 (m, 1H), 3.97 (dd, 2H), 3.41 (t, 2H), 3.33 (s, 3H), 2.57 2.46 (m, 2H), 2.29 (s, 3H), 1.65 (dd, 2H). LC-MS m/z(ESI)= 383.20 [M+1]
Example 3 7-Methyl-2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-9-(tetrahydro-2H-pyran-4 yl)-7,9-dihydro-8H-purin-8-one (compound 3)
F3 C NN N >= I:XN>N H
0 Compound 3
N N F3 C N-N FNC -O CI >zO F3C + H NH 2
0 0 3a Intermediate 1 Compound 3 The title compound 7-methyl-2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 3 (white solid, 50 mg, 21.5% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.62 (s, 1H), 8.12 (s, 1H), 8.00 (d, 1H), 7.53 (s, 1H), 7.47 (d, 1H), 4.47-4.38 (m, 1H), 3.97 (dd, 2H), 3.42 (t, 2H), 3.31 (s, 3H), 2.57-2.40 (m, 2H), 2.35 (s, 3H), 1.69-1.65 (m, 2H). LC-MS m/z(ESI) = 408.20 [M+1]
Example 4 2-((4-Chloro-2-methylphenyl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9 dihydro-8H-purin-8-one (compound 4)
CI N N H
0 Compound 4
N I == !' N I
CI + CI N C N N NH2
0 0 4a Intermediate 1 Compound 4
The title compound 2-((4-chloro-2-methylphenyl)amino)-7-methyl-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-onecompound 4 (white solid, 50 mg, 18.94
% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.49 (s, 1H), 8.04 (s, 1H), 7.60 (d, 1H), 7.26 (d, 1H), 7.17 (dd, 1H), 4.43-4.35 (m, 1H), 3.96 (dd, 2H), 3.40 (t, 2H), 3.29 (s, 3H), 2.50-2.44 (m, 2H), 2.23 (s, 3H), 1.64 (dd, 2H). LC-MS m/z(ESI)= 374.10 [M+1]
Example 5 3-Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzonitrile (compound 5)
NC a N N N >= ~ >N H
0 Compound 5
NC + CI N O NC N N
NH2 H
0 0 5a Intermediate 1 Compound 5 The title compound 3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzonitrile compound 5 (white solid, 50 mg, 18.13 %
yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.68 (s, 1H), 8.15 (s, 1H), 8.06 (d, 1H), 7.63 (s, 1H), 7.57 (dd, 1H), 4.46-4.38 (m, 1H), 3.97 (dd, 2H), 3.42 (t, 2H), 3.32 (s, 3H), 2.55-2.45 (m, 2H), 2.32 (s, 3H), 1.66 (dd, 2H). LC-MS m/z(ESI)= 365.20 [M+1]
Example 6 3-Methyl-4-((7-(methyl-d3)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzonitrile (compound 6)
CD, NC N N N-o NNAj'N N H
0 Compound 6 H CD 3 CD,
CI N S N Step 2 NCN N N o H
0 0o 1D 6a Compound 6 Step 1: 2-Chloro-7-(methyl-d3)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (6a)
2-Chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1D (1.0 g, 3.93 mmol) was dissolved in dimethyl sulfoxide (20 mL), and cesium carbonate (2.03 g, 7.86 mmol) was added at room temperature, followed by addition of deuterated iodomethane (0.45 g, 3.93 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 2 h and monitored by TLC. After the reaction was completed, 5 mL of water was added and the resulting mixture was extracted 3 times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo, and a solid was precipitated. The mixture was filtered and purified by medium pressure preparative liquid chromatography, the filtrate was concentrated under reduced pressure to obtain the title compound 2-chloro-7-(methyl-d3)-9-(tetrahydro 2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 6a (white solid, 0.36 g, 34.66% yield). 1HNMR (400 MHz DMSO) 6 8.36 (s, 1H), 4.50-4.41 (m, 1H), 3.99-3.95 (m, 2H), 3.48
3.42 (m, 2H), 2.47-2.38 (m, 2H), 1.70-1.66 (m, 2H). LC-MS m/z(ESI) = 272.10 [M+1]
Step 2: 3-Methyl-4-((7-(methyl-d3)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzonitrile (compound 6)
4-Amino-3-methylbenzonitrile 5a (200 mg, 1.51 mmol), 2-chloro-7-(methyl-d3)-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 6a (330 mg, 1.21 mmol), cesium carbonate (980 mg, 3.02 mmol), and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (140 mg, 0.15 mmol) were dissolved in dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 100 °C for 4 h and monitored by TLC. After the reaction was completed the mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to obtain compound 3 methyl-4-((7-(methyl-d3)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)benzonitrile compound 6 (light gray solid, 99 mg, 17.56 % yield). 1H NMR (400 MHz, Chloroform-d) 6 8.55 (d, 1H), 7.92 (s, 1H), 7.55 (dd, 1H), 7.46 (d, 1H), 7.09 (s, 1H), 4.60-4.50 (m, 1H), 4.14 (dd, 2H), 3.60-3.48 (m, 2H), 2.83-2.67 (m, 2H), 2.39 (s, 3H), 1.80-1.70 (m, 2H).
LC-MS m/z(ESI) = 368.20 [M+1]
Example 7 2-((4-Fluoro-2-methylphenyl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9 dihydro-8H-purin-8-one (compound 7)
F FX N'j N- I N =O H
0 Compound 7
N N F_ NN
F + CI NC O F NAN O NH2 H
7a Intermediate 1 0 Compound 7
The title compound 2-((4-fluoro-2-methylphenyl)amino)-7-methyl-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 7 (white solid, 50 mg, 17.51
% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 68.45 (s, 1H), 7.99 (s, 1H), 7.46-7.43 (m, 1H), 7.05 (dd, 1H), 6.98-6.93 (m, 1H), 4.41-4.33 (m, 1H), 3.97-3.93 (dd, 2H), 3.4 (t, 2H), 3.27 (s, 3H), 2.54-2.44 (m,2H), 2.2 (s, 3H), 1.63 (dd, 2H). LC-MS m/z(ESI) = 358.20 [M+1]
Example 8 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzonitrile (compound 8)
NCC NNI N >=
F NjN N H
0 Compound 8
N NC NNNN NC +CIN FN N N F - INH 2 H
2A Intermediate 1 Compound 8
The title compound 2-fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzonitrile compound 8 (white solid, 18.1 mg, 10.3 % yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 68.79 (s, 1H), 8.25 (s, 1H), 8.22 (d, 1H), 7.66 (d, 1H), 4.48-4.41 (m, 1H), 3.97 (dd, 2H), 3.43 (t, 2H), 3.32 (s, 3H), 2.57-2.50 (m, 2H), 2.32(s, 3H), 1.68(dd, 2H). LC-MS m/z(ESI)= 383.10 [M+1]
Example 9
2-((6-Methoxy-4-methylpyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl) 7,9-dihydro-8H-purin-8-one (compound 9)
0 N N H
0
Compound 9
N N N~ ' "- N
O A + CI "-I:>=0 NN N~ j1- >szN N N NH b H NH2
0 0
9a Intermediate 1 Compound 9
The title compound 2-((6-methoxy-4-methylpyridin-3-yl)amino)-7-methyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 9 (white solid, 60 mg, 15.2 % yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.49 (s, 1H), 8.04 (s, 1H), 7.95 (s, 1H), 6.70 (s, 1H), 4.41-4.33 (m, 1H), 3.95 (dd, 2H), 3.81 (s, 3H), 3.40 (t, 2H), 3.27 (s, 3H), 2.54-2.44 (m, 2H), 2.15 (s, 3H), 1.63(dd, 2H). LC-MS m/z(ESI) = 371.20 [M+1]
Example 10 3-Chloro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzonitrile (compound 10)
NC CI N
' a NN N H
Compound 10 0I NNC CIN N NC CI + CI N O N CN O NH2 H
10a Intermediate 1 0 Compound 10 0
The title compound 3-chloro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzonitrile compound 10 (white solid, 6.4 mg, 7.8 %
yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz DMSO) 68.61 (s, 1H), 8.44 (d, 1H), 8.23 (s, 1H), 8.05 (d, 1H), 7.77 (d, 1H), 4.47-4.41 (m, 1H), 3.97 (dd, 2H), 3.44 (t, 2H), 1.67 (d, 2H). LC-MS m/z(ESI)= 385.10 [M+1]
Example 11 3-Methyl-4-[(9-(oxa-4-yl)-8-oxo-8,9-dihydro-7H-purin-2-yl)amino]benzonitrile (compound 11)
H NC Na N~ N>=
-NN N
0 Compound11 H H
N NC N N~ nN= NC +~ CI O C N ON N N H NH 2 H 0 0 5a ID Compound11 4-Amino-3-methylbenzonitrile 5a (200 mg, 1.51 mmol), 2-chloro-9-(oxa-4-yl)-8,9 dihydro-7H-purin-8-one 1D (380 mg, 1.51 mmol), cesium carbonate (980 mg, 3.02 mmol), and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (140 mg, 0.15 mmol) were dissolved in dioxane (10 mL), followed by nitrogen purging. The mixture was stirred under nitrogen at 100 °C for 4 h and monitored by TLC. Afterthe reaction was completed. The mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 100/1) to give obtain the title compound 3-methyl 4-[(9-(oxa-4-yl)-8-oxo-8,9-dihydro-7H-purin-2-yl)amino]benzonitrile compound 11 (white solid, 120 mg, 22.68 % yield). 1H NMR (400 MHz, DMSO-d6) 6 11.09 (s, 1H), 8.60 (s, 1H), 8.08 (d, 1H), 7.96 (s, 1H), 7.62 (d, 1H), 7.57 (dd, 1H), 4.42-4.35 (m, 1H), 3.98 (dd, 2H), 3.41 (t, 2H), 2.32 (s, 3H), 1.64 (dd, 2H). LC-MS m/z(ESI)= 351.10 [M+1]
Example 12 2-Fluoro-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzonitrile (compound 12)
F NC_, NI N >=
NC N N H
0 Compound 12 F IF N NCI NN N NC + CI N Nch O C N) NH 2 H
0 0 12a Intermediate 1 Compound 12
The title compound 2-fluoro-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzonitrile compound 12 (pale yellow solid, 25 mg, 7% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 9.00 (s, 1H), 8.20-8.17 (m, 1H), 7.91 (d, 1H), 7.63 (t, 1H), 4.47-4.39 (m, 1H), 3.97 (dd, 2H), 3.45-3.38 (m, 2H), 2.55-2.45 (m, 2H), 2.22 (d, 3H), 1.67 (dd, 2H).
LC-MS m/z(ESI)= 383.20 [M+1]
Example 13 4-((7-Methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino) 3-(trifluoromethyl)benzonitrile (compound 13)
INC CF 3N
N H
0 Compound 13 CIF3 NCIC CF 3 NC CF 3 + CI N 1N->NN " C N/\= N N O +CI' n N>A- > NH2 H
0 0 13a Intermediate 1 Compound 13
The title compound 4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)-3-(trifluoromethyl)benzonitrile compound 13 (pale yellow solid, 35 mg, 11% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz CDC ) 6 8.77 (d, 1H), 8.94-8.80 (m, 4H), 4.59-4.53 (m, 1H), 4.14 3 (dd, 2H), 3.54 (t, 2H), 2.74-2.63 (m, 2H), 1.74 (d, 2H). LC-MS m/z(ESI) = 419.20 [M+1]
Example 14 3-ethyl-4-((7-Methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzonitrile (compound 14)
NC -0N
~-&NN N H
Compound 14 0
NC + CI N - N NAN N
NH 2 H 0 0 14a Intermediate 1 Compound 14
The title compound 3-ethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzonitrile compound 14 (pale yellow solid, 27 mg, % yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.71 (s, 1H), 8.13 (d, 1H), 8.00 (d, 1H), 7.61-7.57 (m, 2H), 4.46-4.38 (m, 1H), 3.97 (dd, 2H), 3.45-3.38 (m, 2H), 2.74 (q, 2H), 1.68-1.64 (d, 2H), 1.15 (t, 3H). LC-MS m/z(ESI)= 379.20 [M+1]
Example 15
4-Methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzonitrile (compound 15)
NC N N H
0 Compound 15
N N N==o N>= + CIN O NC N N NCIO .NC NC NH 2 H 0o 0 15a Intermediate 1 Compound 15 The title compound 4-methyl-3-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzonitrile compound 15 (pale yellow solid, 27 mg, % yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz DMSO) 6 8.67 (s, 1H), 8.14 (d, 2H), 7.39 (d, 2H), 4.41-4.37 (m, 1H), 3.45-3.38 (m, 2H), 3.31 (s, 3H), 2.51-2.47 (m, 2H), 2.35 (s, 3H), 1.70-1.66 (m, 2H). LC-MS m/z(ESI)= 365.20 [M+1]
Example 16 3-Chloro-5-fluoro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 16)
0
H2 N ON N N' NAN N F H
0 Compound 16 0
0 CIN 0 H2 N C1 N" N>= H2 N + CI N N N N N NH 2 F IF 0 0 16a Intermediate 1 Compound 16 The title compound 3-chloro-5-fluoro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 16 (pale yellow solid, 20 mg, 6% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 610.62 (s, 1H), 8.33 (t, 1H), 7.82 (t, 1H), 7.69 (dd, 1H), 4.47-4.39 (m, 1H), 3.96 (dd, 2H), 3.45-3.39 (m, 1H), 3.36 (s, 3H), 2.60-2.53 (m, 2H), 1.64 (dd, 2H). LC-MS m/z(ESI) = 421.20 [M+1]
Example 17 4-Methyl-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)picolinonitrile (compound 17)
NC N NO N '11NAN N H
0 Compound 17
NN N N-N O= NH- CI N O>~ N NCN NC N NH H b 0 0 17a Intermediate 1 Compound 17
The title compound 4-methyl-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)picolinonitrile compound 17 (white solid, 44 mg, 29.3% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz DMSO) 69.12 (s, 1H), 9.05 (s, 1H), 8.17 (s, 1H), 7.87 (s, 1H), 4.46-4.39(m, 1H), 3.97 (dd, 2H), 3.41 (t, 2H), 3.32 (s, 3H), 2.34 (s, 3H), 1.67 (d, 2 H). LC-MS m/z(ESI)= 366.10 [M+1]
Example 18 4-Methyl-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)picolinamide (compound 18)
0
H2 N N N N lj NAN N H
0 Compound 18 0 0 N N H2N N-N H, CI N N> N N H N N
0 0 18a Intermediate 1 Compound 18 The title compound 4-methyl-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)picolinamide compound 18 (white solid, 40 mg, 28.1% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 9.04 (s, 1H), 8.89 (s, 1H), 8.12 (s, 1H), 8.05 (s, 1H), 7.95
(s, 1H), 7.57 (s, 1H), 4.44-4.38 (m, 1H), 3.97 (dd, 2H), 3.44 (t, 2H), 3.31 (s, 3H), 2.36 (s, 3H), 1.67 (d, 2H). LC-MS m/z(ESI)= 384.20 [M+1]
Example 19 2-((5-Methoxy-3-methylpyridin-2-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl) 7,9-dihydro-8H-purin-8-one (compound 19)
0 N- N>= N N N N H
0 Compound 19
N N N N
CI N N NN O H N NH 2 H 0 0 198 Intermediate 1 Compound 19 The title compound 2-((5-Methoxy-3-methylpyridin-2yl)-amino)-7-methyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-onecompound 19 (white solid, 12 mg, 9.6% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 68.73 (s, 1H), 7.96 (s, 1H), 7.90 (d, 1H), 7.28 (d, 1H), 4.38-4.31 (m, 1H), 3.94 (dd, 2H), 3.81 (s, 3H), 3.39 (t, 2H), 3.27 (s, 3H), 2.50-2.42 (m, 2H), 2.15 (s, 3H), 1.61 (dd, 2H). LC-MS m/z(ESI) = 371.20 [M+1]
Example 20 -Methyl-6-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)nicotinonitrile (compound 20)
NC, N N
N "NA'11N N H
0 Compound 20
N N NC NC aC + CI N >=0 NN N A11 N N > N NH2,
0 0 20a Intermediate 1 Compound 20 The title compound 5-methyl-6-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)nicotinonitrile compound 20 (white solid, 85 mg, 20.4% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 69.50 (s, 1H), 8.49 (d, 1H), 8.21 (s, 1H), 7.98 (d, 1H), 4.44-4.36 (m, 1H), 3.96 (dd, 2H), 3.41 (t, 2H), 3.34 (s, 3H), 2.56-2.45 (m, 2H), 2.24 (s, 3H), 1.65 (dd, 2H). LC-MS m/z(ESI) = 366.20 [M+1]
Example 21 2-((6-Methoxy-2-methylpyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl) 7,9-dihydro-8H-purin-8-one (compound 21)
N N H
0 Compound 21
N N - N N ON I11 , + CI N O N N>= I H NH 2 H 0 0
21a Intermediate 1 Compound 21
The title compound 2-((6-Methoxy-2-methylpyridin-3-yl)amino)-7-methyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-onecompound 21 (white solid, 100 mg, 62.17% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 6 8.50 (s, 1H), 7.97 (s, 1H), 7.65 (d, 1H), 6.61 (d, 1H), 4.41-4.32(m, 1H), 3.95 (dd, 2H), 3.81 (s, 3H), 3.39 (t, 2H), 3.27 (s, 3H), 2.51-2.45 (m, 2H), 2.31 (s, 3H), 1.63 (dd, 2H). LC-MS m/z(ESI) = 371.20 [M+1]
Example 22 -Methyl-4-[(7-methyl-9-(oxa-4-yl)-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]thiophene-2-carboxylate (compound 22)
IN -0 N N N N)= N
H N
0 Compound 22
OI+ C NN N N -o NH2 H
0 0 22a Intermediate 1 Compound 22 The title compound 5-methyl-4-[(7-methyl-9-(oxa-4-yl)-8-oxo-8,9-dihydro-7H-purin 2-yl)amino]thiophene-2-carboxylate compound 22 (pale yellow solid, 50 mg, 11.09% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz, DMSO-d6) 6 8.90 (s, 1H), 8.08 (s, 1H), 8.05 (s, 1H), 4.40-4.35
(m, 1H), 3.96 (dd, 2H), 3.79 (s, 3H), 3.41 (t, 2H), 3.29 (s, 3H), 2.37 (s, 3H), 1.67-1.64 (m, 2H). LC-MS m/z(ESI) = 404.10 [M+1]
Example 23 7-Methyl-2-((4-methylpyrimidin-5-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9 dihydro-8H-purin-8-one (compound 23)
NI KN N'_ N )= N N 'IN " N H
0
Compound 23
NCI' NN N N
NH 2 H
0 0 23a Intermediate 1 Compound 23
The title compound 7-methyl-2-((4-methylpyrimidin-5-yl)amino)-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 23 (white solid, 50 mg, 21.5% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 68.92 (s, 1H), 8.90 (s, 1H), 8.73 (s, 1H), 8.0 (s, 1H), 4.44
4.36 (m, 1H), 3.96 (dd, 2H), 3.40 (t, 2H), 3.30 (s, 3H), 2.53-2.46 (m, 2H), 2.45 (s, 3H), 1.65 (dd, 2H). LC-MS m/z(ESI) = 342.20 [M+1]
Example 24 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 24)
0
HN _' N"'- N0 F NN N H
Compound 24 0 I 0 0 NN
F N N F NH2 H 0b Intermediate 2 Intermediate 1 Compound 24 0
The title compound 2-fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 24 (white solid, 9.5 mg, 8.6% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 68.54 (s, 1H), 8.18 (s, 1H), 7.89 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.39 (s, 1H), 4.48-4.40 (m, 1H), 3.98 (dd, 2H), 3.42 (t, 2H), 2.58-2.53 (m, 2H), 2.29 (s, 3H),1.67 (dd, 2H). LC-MS m/z(ESI)= 401.20 [M+1]
Example 25 3-(Methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzonitrile (compound 25)
C NC N / N
N N1 N N H
0 Compound 25 CID, NC CD 3 I1:-' + CI N ON N N NH 2 H
0 0
25a Intermediate I Compound 25
The title compound 3-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl) 8,9-dihydro-7H-purin-2-yl)amino)benzonitrile compound 25 (pale yellow solid, 40 mg, 29.4% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz, DMSO-d )6 8.68 (s, 1H), 8.16 (s, 1H), 8.07 (d, 1H), 7.63 (d, 1H), 7.59-7.57 (m, 1H), 4.46-4.39 (m, 1H), 3.99-3.97 (m, 2H), 3.42 (t, 2H), 3.32 (s, 3H), 2.57-2.50 (m, 2H), 1.69-1.65 (m, 2H). LC-MS m/z(ESI)= 368.20 [M+1]
Example 26 3-Chloro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide (compound 26)
0
H 2N CI N O -a N' N N H
0 Compound 26 0
NC CI Step1 0 H 2N C IN N Stp1C1 Step 2 N ~ N/>0 H2NN NH2 H NH2 IaNH 2 H b 26a 26b Compound 26 Step 1: 4-Amino-3-chlorobenzamide (26b)
4-Amino-3-chlorobenzonitrile 26a (2 g, 13 mmol) and potassium carbonate (271 mg, 1.9 mmol) were dissolved in 6 mL of dimethyl sulfoxide, and 1.78 mL of hydrogen peroxide was added in an ice bath. The reaction mixture was then gradually heated to °C and stirred for 2 h. The reaction was completed as monitored by TLC. 10 mL of water was added to the reaction mixture, then a white solid was precipitated. The mixture was filtered, and the filter cake was collected and dried to obtain the title compound 4-amino-3-chlorobenzamide 26b (white solid, 1.0 g, 48% yield). 1H NMR (400 MHz DMSO) 6 7.75 (s, 1H), 7.57 (d, 1H), 7.03 (s, 1H), 7.60 (d, 1H), 5.87 (s, 2H). LC-MS m/z(ESI)= 171.10 [M+1]
Step 2: 3-Chloro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide (compound 26)
The title compound 3-chloro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide compound 26 (white solid, 9.4 mg, 8.3% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 68.39 (s, 1H), 8.27 (d, 1H), 8.19 (s, 1H), 7.99 (d, 1H), 7.97 (s, 1H), 7.83 (dd, 1H), 7.36 (s, 1H), 4.44-4.34 (m, 1H), 3.98 (dd, 2H), 3.44 (t, 2H), 2.55-2.45 (m, 5H), 1.68(dd, 2H). LC-MS m/z(ESI) = 403.10 [M+1]
Example 27 N,3-Dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 27)
0 N
H
0 Compound 27 0 N N HOOCa. N~ N X N HOOC + CI -0 NStep 1 HOOC N --O Step 2 H N H H NH 2 H H
0 0 0 27a Intermediate 1 27b Compound 27
Step 1: 3-Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzoic acid (27b)
3-Methyl-4-aminobenzoic acid 27a (2.0 g, 13.4 mmol), 2-chloro-7-methyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one intermediate 1 (3.0 g, 11.2 mmol), cesium carbonate (7.3 g, 22.4 mmol) and catalyst Brettphos-G3-Pd (CAS: 1470372-59-8) methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri i-propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (1.0 g, 1.12 mmol) were added to a 50 mL flask, followed by addition of 30 mL of 1,4-dioxane. After being purged with nitrogen three times, the reaction system was heated to reflux for 3 h. The reaction was completed as monitored by TLC. The reaction mixture was diluted with dichloromethane and filtered through celite, and water was added to the filtrate. The resulting mixture was concentrated under reduced pressure to remove a small amount of organic solvent, and a large amounts of yellow solid was precipitated. The solid was collected by filtration and slurried with ethanol, and the slurry was filtered and dried by rotary evaporation to obtain the title compound 3-methyl-4-((7-methyl-8-oxo-9 (tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzoic acid 27b (pale yellow solid, 2.15 g, 50% yield). LC-MS m/z(ESI) = 384.20 [M+1]
Step 2: N,3-Dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 27)
3-Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzoic acid 27b (250 mg, 0.65 mmol), 1-(3-dimethylaminopropyl)-3 ethylcarbodiimide hydrochloride (187.2 mg, 0.98 mmol), 1-hydroxybenzotriazole (132.4 mg, 0.98 mmol), and triethylamine (333 mg, 3.3 mmol) were dissolved in 2 mL of dichloromethane. The reaction mixture was heated to 40 °C and stirred for 1 h, and then methylamine hydrochloride (220 mg, 3.3 mmol) was added. The reaction system was stirred at 40 °C for 3 h. The reaction was completed as monitored by TLC. Water was added to the mixture and extracted three times with an appropriate amount of dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate. The reaction mixture was filtered and concentrated to give a crude product, which was then slurried with methanol. The slurry was filtered, and the solid was dried under reduced pressure to obtain the title compound N,3-dimethyl-4-((7 methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide compound 27 (white solid, 21 mg, 8.1% yield). 'H NMR (400 MHz DMSO) 68.48 (s, 1H), 8.27 (q, 1H), 8.10 (s, 1H), 7.82 (d, 1H), 7.67 (d, 1H), 7.62 (dd, 1H), 4.45-4.37 (m, 1H), 3.97 (dd, 2H), 3.42 (t, 2H), 3.31 (s, 3H), 2.76 (d, 3H), 2.57-2.46 (m, 2H), 2.30 (s, 3H), 1.66 (dd, 2H). LC-MS m/z(ESI)= 397.20 [M+1]
Example 28 N,N,3-Trimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 28)
0 N
N N - H
0 Compound 28 0 HOOCa N O N N N N-a H H
0 0 Compound 28 27b The title compound N, N,3-trimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 28 (white solid, 65 mg, 24% yield) was prepared according to the synthesis method of compound 27. 1H NMR (400 MHz DMSO) 68.49 (s, 1H), 8.07 (s, 1H), 7.72 (d, 1H), 7.24 (d, 1H), 7.19 (dd, 2H), 4.40-4.36 (m, 1H), 3.95 (dd, 2H), 3.43-3.37 (m, 2H), 3.30 (s, 3H), 2.96 (s, 6H), 2.55-2.45 (m, 2H), 2.27 (s, 3H), 1.65 (dd, 2H). LC-MS m/z(ESI) = 411.20 [M+1]
Example 29 N-Ethyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 29)
0I N
N N -N H
0 Compound 29
HOOC H0C. NN- NNN NO NN H NN N H H
0 0 Compound 29 27b The title compound N-ethyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 29 (white solid, 67 mg, 24% yield) was prepared according to the synthesis method of compound 27. 1H NMR (400 MHz, DMSO-d6) 68.47(s, 1H), 8.29 (t, 1H), 8.10 (s, 1H), 7.83 (d, 1H), 7.68 (d, 1H), 7.63 (dd, 1H), 4.45-4.37 (m, 1H), 3.97 (dd, 2H), 3.45-3.38 (m, 2H), 3.33 3.23 (m, 5H), 2.57-2.46 (m, 2H), 2.30 (s, 3H), 1.68-1.64 (m, 2H), 1.11 (t, 3H). LC-MS m/z(ESI) = 411.20 [M+1]
Example 30 N-Isopropyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 30)
0I N N N H) N N N )= H
0 Compound 30 / 0/ HOOC. N N -ONN O N N1 N H ' N N'N H H
o 0 Compound 30 27b The title compound N-isopropyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 30 (white solid, 67 mg, 23% yield) was prepared according to the synthesis method of compound 27. 1H NMR (400 MHz, DMSO-d6) 6 8.47 (s, 1H), 8.10 (s, 1H), 8.03 (d, 1H), 7.82 (d, 1H), 7.69 (d, 1H), 7.63 (dd, 1H), 4.44-4.38 (m, 1H), 4.11-4.06 (m 1H), 3.97 (dd, 2H), 3.42 (t, 2H), 3.30 (s, 3H), 2.54-2.46 (m, 2H), 2.30 (s, 3H), 1.66 (dd, 2H), 1.16 (d, 6H). LC-MS m/z(ESI) = 425.20 [M+1]
Example 31 N-Cyclopropyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide (compound 31)
AlN0 N' N HH HOOC NN ON N H
0 H Compound 31
NN N ~ N
H b-H
0 0 27b Compound 31
The title compound N-cyclopropyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 31 (white solid, 135 mg, 46% yield) was prepared according to the synthesis method of compound 27. 'H NMR (400 MHz, DMSO-d6) 8.46 (s, 1H), 8.30 (d, 1H), 8.10 (s, 1H), 7.81 (d, 1H), 7.68 (d, 1H), 7.62 (dd, 1H), 4.45-4.37 (m, 1H), 3.97 (dd, 2H), 3.42 (t, 2H), 3.31 (s, 3H), 2.86-2.80 (m, 1H), 2.57-2.46 (m, 2H), 2.30 (s, 3H), 1.67 (dd, 2H), 0.69-0.65 (m, 2H), 0.59-0.55 (m, 2H). LC-MS m/z(ESI) = 423.20 [M+1]
Example 32 N-Isobutyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 32)
0 Nr N
H
0 Compound 32 0 HOOC N N N N O H H
0 0 Compound 32 27b The title compound N-isobutyl-3-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran 4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 32 (white solid, 21 mg, 8% yield) was prepared according to the synthesis method of compound 27. 1H NMR (400 MHz, DMSO-d6) 68.48 (s, 1H), 8.28 (t, 1H), 8.10 (s, 1H), 7.82 (d, 1H), 7.70 (d, 1H), 7.64 (dd, 1H), 4.45-4.37 (m, 1H), 4.97 (dd, 2H), 3.42 (q, 2H), 3.30 (s, 3H), 3.70 (dd, 2H), 2.57-2.47 (m, 2H), 2.30 (s, 3H), 1.87-1.80 (m, 1H), 1.67 (dd, 2H), 0.92-0.86 (m, 6H). LC-MS m/z(ESI) = 439.30 [M+1]
Example 33 7-Methyl-2-((2-methyl-4-(morpholine-4-carbonyl)phenyl)amino)-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 33)
0 N N N
H
0 Compound 33 0 HOOCa. N N-a N N Q N>=j > NNNN N N H H
0 0 Compound 33 27b The title compound 7-methyl-2-((2-methyl-4-(morpholine-4-carbonyl)phenyl)amino) 9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 33 (white solid, 21 mg, 7.1% yield) was prepared according to the synthesis method of compound 27. 1H NMR (400 MHz DMSO) 6 8.52 (s, 1H), 8.07 (s, 1H), 7.73 (d, 1H), 7.25 (d, 1H), 7.19 (dd, 1H), 4.42-4.36 (m, 1H), 3.94 (dd, 2H), 3.59-3.40 (m, 9H), 3.29 (s, 3H), 2.27 (s, 3H), 1.67-1.62 (m, 2H). LC-MS m/z(ESI) = 453.20 [M+1]
Example 34 2-((4-(3-Hydroxypyrrolidine-1-carbonyl)-2-methylphenyl)amino)-7-methyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 34)
0
HO- N ~ NN N H
0 Compound 34 0 HOOCX N HO N N N == N N N H H
0 0 Compound 34 27b The title compound 2-((4-(3-hydroxypyrrolidine-1-carbonyl)-2-methylphenyl)amino) 7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 34 (white solid, 65 mg, 25% yield) was prepared according to the synthesis method of compound 27. H NMR (400 MHz DMSO) 6 8.49 (s, 1H), 8.08 (s, 1H), 7.75 (d, 1H), 7.29-7.46 (m, 2H), 5.01-4.91 (m, 1H), 4.43-4.22 (m, 2H), 4.03-3.94 (m, 2H), 3.64-3.37 (m, 5H), 3.30 (s,3H), 2.55-2.49 (m, 2H), 2.28 (s, 3H), 2.05-1.79 (m, 2H), 1.65 (d, 2H). LC-MS m/z(ESI) = 453.20 [M+1]
Example 35
3-Methyl-N-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide (compound 35)
0
N NN H
Compound 35 0 HOOC N N DC - N N H u' >= N N =' NN N N H H
0 0 Compound 35 27b The title compound 3-methyl-N-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 35 (white solid, mg, 22% yield) was prepared according to the synthesis method of compound 27. 1H NMR (400 MHz DMSO) 6 8.48 (s, 1H), 8.24 (s, 1H), 8.01 (s, 1H), 7.82 (d, 1H), 7.67 (d, 1H), 7.62 (dd, 1H), 4.44-4.38 (m, 1H), 3.97 (dd, 2H), 3.44-3.38 (m, 2H), 3.30 (s,3H), 2.30 (s, 3H), 1.68-1.64 (m, 2H). LC-MS m/z(ESI)= 400.20 [M+1]
Example 36 2-Fluoro-N,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide (compound 36)
0/
HN N H
Compound 36 0
0 0 N0N:_ HO Step1 2 H O 1.HF a N N N F NH 2 F NH 2 H
36a 36b Compound 36 O Step 1: 4-Amino-2-fluoro-N,5-dimethylbenzamide (36b)
4-Amino-2-fluoro-5-methylbenzoic acid 36a (200 mg, 1.18 mmol) was dissolved in thionyl chloride (10 mL). The mixture was heated to reflux for 1 h and concentrated under reduced pressure to evaporate the thionyl chloride, and 10 mL of toluene was added to remove excess thionyl chloride. The residue was dissolved in 10 mL of dichloromethane, and the resulting solution was added dropwise to a solution of potassium carbonate (820 mg, 5.9 mmol) and methylamine hydrochloride (399 mg, 5.90 mmol) in dichloromethane (10 mL). The reaction mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove dichloromethane, and the residue was purified by column chromatography (dichloromethane/methanol (v/v) = 20/1) to obtain the title compound 4-amino-2 fluoro-N,5-dimethylbenzamide 36b (pale yellow solid, 0.2 g, 92.84% yield). 1H NMR (400 MHz, DMSO-d) 6 7.52 (t, 1H), 7.32 (d, 1H), 6.32(d, 1H), 5.65 (s, 2H), 2.72 (d, 3H), 2.01 (s, 3H). LC-MS m/z(ESI) = 183.10 [M+1]
Step 2: 2-Fluoro-N,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide (compound 36)
The title compound 2-fluoro-N,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 36 (white solid, 17 mg, 14.95% yield) was prepared according to the synthesis method of compound 1. 1H NMR (401 MHz, DMSO-d) 68.51 (s, 1H), 8.17 (s, 1H), 7.95 (s, 1H), 7.89 (d, 1H), 7.49 (d, 1H), 4.43 (t, 1H), 3.98 (d, 2H), 3.32 (s, 3H), 2.76 (d, 3H), 2.29 (s, 3H), 1.68 (d, 2H). LC-MS m/z(ESI) = 415.20 [M+1]
Example 37 2-Fluoro-5-methyl-N-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl) 8,9-dihydro-7H-purin-2-yl)amino)benzamide (compound 37)
O D N NN H
Compound 37 0
0 Step H Step2 D0N N
H> te F F NH NN:: N> N F:a ' H Fa NH 2
36a 37a Compound 37 0
Step 1: 4-Amino-2-fluoro-5-methyl-N-(methyl-d3)benzamide (37a)
4-Amino-2-fluoro-5-methylbenzoic acid 36a (200 mg, 1.18 mmol) was dissolved in thionyl chloride (10 mL). The mixture was heated at reflux for 1 h and concentrated to evaporate the thionyl chloride, and 10 mL of toluene was added to remove excess thionyl chloride. The residue was dissolved in 10 mL of dichloromethane, and the resulting solution was added dropwise to a solution of potassium carbonate (820 mg, 5.9 mmol) and deuterated methylamine (200 mg, 5.90 mmol) in dichloromethane (10 mL). The reaction mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by column chromatography (dichloromethane/methanol (v/v) = 20/1) to obtain the title compound 4-amino-2 fluoro-5-methyl-N-(methyl-d3)benzamide 37a (pale yellow solid, 0.2 g, 91.51% yield).
'H NMR (400 MHz, DMSO-d) 6 7.54-7.46 (m, 1H), 7.32 (d, 1H), 6.32 (d, 1H), 5.65 (s, 2H), 2.01 (s, 3H). LC-MS m/z(ESI) = 186.10 [M+1]
Step 2: 2-Fluoro-5-methyl-N-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl) 8,9-dihydro-7H-purin-2-yl)amino)benzamide (compound 37)
The title compound 2-fluoro-5-methyl-N-(methyl-d3)-4-((7-methyl-8-oxo-9 (tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 37 (white solid, 15 mg, 13.31% yield) was prepared according to the synthesis method of compound 1. 1H NMR (401 MHz, DMSO-d) 68.51 (s, 1H), 8.17 (s, 1H), 7.92-7.87 (m, 2H), 7.49
(d, 1H), 4.43 (t, 1H), 3.97 (d, 2H), 3.32 (s, 3H), 2.28 (s, 3H), 1.68 (d, 2H). LC-MS m/z(ESI) = 418.10 [M+1]
Example 38 2-((6-(Methoxy-d3)-4-methylpyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran 4-yl)-7,9-dihydro-8H-purin-8-one (compound 38)
D+D
O N N N. NA' N N)= H
Compound 38 o
D NH 2 NO2 Step 2 F NO2 Step I F' N D0L 0 'CN D 0 'N'
38a 38b 38c
D D
Step3 D* NN N N N H
Compound 38 o Step 1: 2-(Methoxy-d3)-4-methyl-5-nitropyridine (38b)
2-Fluoro-4-methyl-5-nitropyridine 38a (0.5 g, 3.20 mmol) was dissolved in acetonitrile (10 mL), and potassium carbonate (1.33 g, 9.60 mmol) and deuterated methanol (0.21 g, 6.4 mmol) were added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the solid was washed with 10 mL of acetonitrile. The filtrates were combined and concentrated to get a crude product, which was then purified by column chromatography (petroleum ether/ethyl acetate (v/v) = 10/1) to give the title compound 2-(methoxy-d3)-4-methyl-5-nitropyridine 38b (white solid, 0.4 g, 73.03% yield). 1H NMR (400 MHz, DMSO-d) 6 8.91 (s, 1H), 6.96 (s, 1H), 2.55 (s, 3H).
LC-MS m/z(ESI) = 172.10 [M+1]
Step 2: 6-(Methoxy-d3)-4-methylpyridin-3-amine (38c)
2-(Methoxy-d3)-4-methyl-5-nitropyridine 38b (100 mg, 0.58 mmol) was dissolved in ethanol (20 mL), and iron powder (163.13 mg, 2.92 mmol) and acetic acid (20 tL) were added. The reaction mixture was heated to 90 °C and reacted for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the solid was washed with 20 mL of ethanol and purified by column chromatography (petroleum ether/ethyl acetate (v/v)= 100/1) to get the title compound 6-(methoxy-d3)-4-methylpyridin-3-amine 38c (yellow solid, 50 mg, 61% yield). 1H NMR (400 MHz, DMSO-d) 7.47 (s, 1H), 6.45 (s, 1H), 4.51 (s, 2H), 2.05 (d, 3H). LC-MS m/z(ESI)= 142.10 [M+1]
Step 3: 2-((6-(Methoxy-d3)-4-methylpyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran 4-yl)-7,9-dihydro-8H-purin-8-one (compound 38)
The title compound 2-((6-(methoxy-d3)-4-methylpyridin-3-yl)amino)-7-methyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 38 (white solid, mg, 22.73% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz, Chloroform-d) 68.37 (s, 1H), 7.77 (s, 1H), 6.65 (s, 1H), 4.55-4.45
(m, 1H), 4.10 (dd, 2H), 3.56-3.47 (m, 2H), 3.37 (s, 3H), 2.75-2.68 (m, 2H), 2.27 (s, 3H), 1.68 (dd, 2H). LC-MS m/z(ESI) = 374.20 [M+1]
Example 39 4-Methyl-N-(methyl-d3)-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)picolinamide (compound 39)
D DO D> N NI '~ N H I~N NO H
Compound 39 0 0 D 0 D 0 Step1 D Step 2 DNH HO U" . D> N D N N N H NO 2 NO2 NH
39a 39b 39c
D O Step 3 D >ON N H N N H
Compound 39 0
Step 1: 4-Methyl-N-(methyl-d3)-5-nitropicolinamide (39b)
4-Methyl-5-nitropyridine carboxylic acid 39a (430 mg, 2.3 mmol) and 0-(7 azabenzotriazol-1-yl)-N,N,N,N-tetramethylurea hexafluorophosphate (1.04 g, 2.7 mmol) were dissolved in dichloromethane (10 mL). The resulting solution was stirred at 0 °C for 20 min, and N,N-diisopropylethylamine (11 g, 8.2 mmol) and methane-d3 amine (500 mg, 7.08 mmol) were added. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, 10 mL of water was added to the reaction mixture, and the organic phase was separated, dried over anhydrous sodium sulfate and purified by preparative medium pressure normal-phase chromatography to obtain the title compound 4-methyl-N (methyl-d3)-5-nitropicolinamide 39b (brown solid, 142 mg, 41% yield). 'H NMR (400 MHz DMSO) 6 9.07 (s, 1H), 8.21 (s, 1H), 7.96 (s, 1H), 2.72 (s, 3H). LC-MS m/z(ESI)= 199.20 [M+1]
Step 2: -Amino-4-methyl-N-(methyl-d3)picolinamide (39c)
4-Methyl-N-(methyl-d3)-5-nitronaphthamide 39b (142 mg, 0.71 mmol) was dissolved in 5 mL of acetic acid, and zinc powder was added with stirring at 0 °C. The reaction mixture was gradually warmed to room temperature and stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered and concentrated to obtain the title compound 5-amino-4-methyl-N-(methyl d3)picolinamide 39c (yellow solid, 80 mg, 60% yield). LC-MS m/z(ESI)= 169.20 [M+1]
Step 3: 4-Methyl-N-(methyl-d3)-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)picolinamide (compound 39)
The title compound 4-methyl-N-(methyl-d3)-5-((7-methyl-8-oxo-9-(tetrahydro-2H pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)picolinamide compound 39 (white solid, 16 mg, 21.1% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz DMSO) 6 8.83 (s, 2H), 8.56 (s, 1H), 8.09 (s, 1H),7.87 (s, 1H), 4.44 4.38 (m, 1H), 3.97 (d, 2H), 3.44-3.34 (m, 2H), 3.31(s,3H), 2.33 (s, 3H), 1.67 (d, 2H). LC-MS m/z(ESI)= 401.20 [M+1]
Example 40 7-Methyl-2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-9-(tetrahydro-2H-pyran-4 yl)-7,9-dihydro-8H-purin-8-one (compound 40)
H O / N N
~ ~N N = H
0 Compound 40
H O N N N N ~~N N0 N In N>= H H
0 0 Compound 21 Compound 40
2-((6-Methoxy-2-methylpyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl) 7,9-dihydro-8H-purin-8-one compound 21 (400 mg, 1.08 mmol), lithium chloride (228.88 mg, 5.40 mmol) andp-toluenesulfonic acid (1.03 g, 5.40 mmol) were dissolved in DMF (10 mL), and the mixture was stirred at 120 °C for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to obtain the title compound 7-methyl-2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-9-(tetrahydro 2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 40 (white solid, 200 mg, 51.97 % yield). 'H NMR (400 MHz DMSO) 6 11.53(s,1H), 8.25(s,1H), 7.95 (s,1H), 7.31 (d,1H), 6.12 (d, 1H), 4.38-4.31 (m, 1H), 3.97-3.93 (m, 2H), 3.39 (t, 2H), 3.25 (s, 3H), 2.57-2.42 (m, 2H), 2.07 (s, 3H), 1.64-1.60 (m, 2H). LC-MS m/z(ESI)= 357.20 [M+1]
Example 41 -Methyl-4-[(7-methyl-9-(oxepan-4-yl)-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]thiophene-2-carboxylic acid (compound 41)
X I O S J N~N HO H N 0
Compound 41
0N N == 0 \ I N~ N>=H0 N>IiN N N H H -O HN
b-0 0
Compound 22 Compound 41
-Methyl-4-[(7-methyl-9-(oxetan-4-yl)-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]thiophene-2-carboxylate compound 22 (0.78 g, 1.93 mmol) was dissolved in tetrahydrofuran (10 mL) and water (10 mL), and lithium hydroxide (0.09 g, 3.86 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to about 5 with 6 N hydrochloric acid, and a white solid was precipitated. The reaction mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to provide the title compound 5-methyl-4-[(7-methyl-9-(oxepan-4-yl)-8-oxo-8,9-dihydro-7H-purin 2-yl)amino]thiophene-2-carboxylic acid compound 41 (brown solid, 0.4 g, 53.22% yield). 1H NMR (400 MHz, DMSO-d) 6 12.80 (s, 1H), 8.85 (s, 1H), 8.06 (s, 1H), 7.93 (s, 1H), 4.42-4.34 (m, 1H), 3.96 (dd, 2H), 3.39 (t, 2H), 3.29 (s, 3H), 2.35 (s, 3H), 1.67-1.63 (m, 2H). LC-MS m/z(ESI) = 390.10 [M+1]
Example 42 -Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)thiophene-2-carboxamide (compound 42)
o S N
HN N N N H b 0 Compound 42
0 NN N)=
HO N N H H Hb b 0 0 Compound41 Compound42
-Methyl-4-[(7-methyl-9-(oxepan-4-yl)-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]thiophene-2-carboxylic acid compound 41 (0.3 g, 0.77 mmol) was dissolved in anhydrous dichloromethane (20 mL), and a catalytic amount of NN dimethylformamide was added. The reaction mixture was cooled to 0 °C and stirred for min, and oxalyl dichloride (0.1 mL, 1.16 mmol) was added dropwise to the mixture. The reaction mixture was warmed to room temperature and stirred for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the solid was collected and added to 11 mL of dichloromethane/ammonia (10:1) in an ice bath. The mixture was stirred at room temperature for 1 h and concentrated under reduced pressure to give a crude product, which was then purified by preparative medium pressure liquid chromatography to obtain the title compound 5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl) 8,9-dihydro-7H-purin-2-yl)amino)thiophene-2-carboxamide compound 42 (white solid, 140 mg, 46% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.73 (s, 1H), 8.03 (s, 1H), 7.83 (s, 1H), 7.73 (s, 1H), 7.26 (s, 1H), 4.42-4.34 (m, 1H), 3.95 (dd, 2H), 3.40 (t, 2H), 3.29 (s, 3H), 2.29 (s, 3H), 1.65 (dd, 2H). LC-MS m/z(ESI)= 389.20 [M+1]
Example 43 -Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)thiophene-2-carbonitrile (compound 43)
NC N N H
Compound 43
SN QS N" NNC ~-k~>sz0 H2 N Q N N O -1L -- N H N N H
,b 0
Compound 41 Compound 43
-Methyl-4-[(7-methyl-9-(oxetan-4-yl)-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]thiophene-2-carboxamide compound 41 (80 mg, 0.21 mmol) was dissolved in dichloromethane (10 mL), and pyridine (70 mg, 0.84 mmol) and trifluoroacetic anhydride (130 mg, 0.63 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 0.5 h and monitored by TLC. After completion of the reaction, the reaction mixture was quenched with methanol (10 mL) and concentrated to give a crude product, which was then purified by preparative medium pressure liquid chromatography (30% water/acetonitrile) to obtain the title compound 5-methyl-4-[(7 methyl-9-(-4-yl)-8-oxo-8,9-dihydro-7H-purin-2-yl)amino]thiophene-2-carbonitrile compound 43 (pale yellow solid, 20 mg, 25.71% yield). 'H NMR (400 MHz, DMSO-d) 6 9.02 (s, 1H), 8.08 (s, 1H), 8.07 (s, 1H), 4.43-4.36 (m, 1H), 3.98 (dd, 2H), 3.44-3.38 (m, 2H), 3.29 (s, 3H), 2.38 (s, 3H), 1.67-1.64 (m, 2H). LC-MS m/z(ESI)= 371.10 [M+1]
Example 44 2-Fluoro-5-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide (compound 44)
CD 3 I H 2N C O~
F N<N N H
Compound 44 00 CD 3 /
NC H 2N CD 3 Step 2 H2N C
F N2 FF N N N F~NF NH, H
44a 44b Compound 44 0
Step 1: 4-Amino-2-fluoro-5-(methyl-d3)benzamide (44b)
4-Amino-2-fluoro-5-(methyl-d3)benzonitrile 44a (678 mg, 4.72 mmol) was dissolved in DMSO (2 mL), and hydrogen peroxide (3 mL) was slowly added dropwise with stirring at room temperature. The reaction mixture was stirred for 2 h and monitored by TLC. After completion of the reaction,the reaction mixture was poured into purified water (15 mL), and a large amounts of solid was precipitated. The solid was collected by filtration and dried under reduced pressure to remove the solvent and thus to get the title compound 4-amino-2-fluoro-5-(methyl-d3)benzamide 44b (yellow solid, 609 mg, % yield). 'H NMR (400 MHz, DMSO-d) 67.38 (d, 1H), 7.14 (s, 1H), 6.95 (s, 1H), 6.33 (d, 1H), 5.68 (s, 2H). LC-MS m/z(ESI)= 172.10 [M+1]
Step 2: 2-Fluoro-5-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)benzamide (compound 44)
The title compound 2-fluoro-5-(methyl-d3)-4-((7-methyl-8-oxo-9-(tetrahydro-2H pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 44 (pale yellow solid, 120 mg, 27% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz, DMSO-d) 68.52 (s, 1H), 8.19 (d, 1H), 7.90 (d, 1H), 7.55 (d, 1H), 7.42 (d, 2H), 4.47-4.39 (m, 1H), 4.00-3.96 (m, 2H), 3.44-3.35 (m 2H), 3.33 (s, 3H), 2.57-2.55(m, 2H), 1.71-1.67 (m, 2H). LC-MS m/z(ESI) = 404.20 [M+1]
Example 45 2-Cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzoate (compound 45)
0I N N ON H
Compound 45 0
Step I Br Step 2 N ON N0 NH 2 N Ca N'N NH 2 NC NH 2 H
15a 45a 45b Compound 45 0
Step 1: -Amino-2-bromo-4-methylbenzonitrile (45a)
-Amino-4-methylbenzonitrile 15a (2.5 g, 18.9 mmol) was dissolved in DMF (15 mL), and NBS (3.37 g, 18.9 mmol) was added. The reaction mixture was stirred at room temperature for 0.5 h and monitored by TLC. After completion of the reaction, the reaction mixture was poured into purified water (100 mL), and a large amounts of solid was precipitated. The solid was collected by filtration and dried under reduced pressure to obtain the title compound 5-amino-2-bromo-4-methylbenzonitrile 45a (yellow solid, 3.5 g, 87.5% yield). LC-MS m/z(ESI) = 210.90 [M+1]
Step 2: Methyl 4-amino-2-cyano-5-methylbenzoate (45b)
-Amino-2-bromo-4-methylbenzonitrile 45a (3.5 g, 16.6 mmol) was dissolved in toluene (15 mL) and methanol (15 mL), and 4-dimethylaminopyridine (2.0 g, 16.6 mmol), palladium(II) acetate (186 mg, 0.83 mmol) and (9,9-Dimethyl-9H-xanthene 4,5-diyl)bis(diphenylphosphane) (961 mg, 1.66 mmol) were added. The system was purged 4 times with nitrogen. Dicobalt octacarbonate (2.84 g, 8.3 mmol) was rapidly poured into the reaction mixture, and the resulting mixture was refluxed at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was purified by column chromatography to obtain the title compound methyl 4-amino-2-cyano-5-methylbenzoate 45b (yellow solid, 500 mg, 15.9% yield). LC-MS m/z(ESI)= 191.10 [M+1]
Step 3: 2-Cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzoate (compound 45)
The title compound 2-cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzoate compound 45 (pale yellow solid, 502 mg, 45% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz DMSO) 6 8.92 (s, 1H), 8.55 (s, 1H), 8.25 (s, 1H), 7.94 (s, 1H), 4.45-4.38 (m, 1H), 4.00-3.95 (m, 2H), 3.88 (s, 3H), 3.47-3.42 (m, 2H), 3.33 (s, 3H), 2.67-2.50 (m, 2H), 2.43 (s, 3H), 1.72-1.67 (m, 2H). LC-MS m/z(ESI) = 423.20 [M+1]
Example 46 2-Cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzoic acid (compound 46)
0 HON- N' O
H
Compound 46 o 0
HON NN N N NC ON NN NC N " Nn N>= H H
Compound 45 0 Compound 46
2-Cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzoate compound 45 (200 mg, 0.47 mmol) was added to a mixture of tetrahydrofuran (2 mL) and water (2 mL), and lithium hydroxide monohydrate (94 mg, 2.35 mmol) was added. The resulting mixture was stirred at room temperature overnight. The reaction was monitored by TLC. After completion of the reaction, the pH of the reaction mixture was adjusted to about 5 with concentrated hydrochloric acid in an ice bath and concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography to obtain the title compound 2-cyano -methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzoic acid compound 46 (pale yellow solid, 90 mg, 47% yield). 1H NMR (400 MHz, DMSO-d6) 6 13.39 (s, 1H), 8.88 (s, 1H), 8.50 (s, 1H), 8.24 (d, 1H), 7.93 (s, 1H), 4.48-4.40 (m,, 1H), 4.01-3.97 (m, 2H), 3.48-3.35 (m, 2H), 3.34 (s,
3H), 2.57-2.55 (m, 2H), 2.42 (s, 3H), 1.73-1.68 (m, 2H). LC-MS m/z(ESI) = 409.20 [M+1]
Example 47 2-Cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 47)
0I
H2 N N NO NC ):jN N N H
Compound 47 O 0
HON N NN H2N N N NC N N N>= NC N<N N H H
Compound 46 Compound 47
2-Cyano-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzoic acid compound 46 (75 mg, 0.183 mmol) was added to 2 mL of tetrahydrofuran, then HATU (104 mg, 0.275 mmol) and triethylamine (28 mg, 0.275 mmol) were added. The reaction mixture was stirred at room temperature for 0.5 h, and then 2 mL of ammonia in 1,4-dioxane solution (0.5 M in 1,4-dioxane) was slowly added dropwise. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by column chromatography to obtain the title compound 2-cyano-5-methyl-4-((7-methyl-8-oxo-9 (tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 47 (pale yellow solid, 42 mg, 56% yield). 1H NMR (400 MHz, DMSO-d6) 8.80 (s, 1H), 8.34 (s, 1H), 8.20 (s, 1H), 8.00 (s, 1H), 7.72 (s, 1H), 7.53 (s, 1H), 4.47-4.39 (m, 1H), 4.01-3.96 (m, 2H), 3.49-3.38 (m, 2H), 3.32 (s, 3H), 2.52-2.50(m, 2H), 2.39 (s, 3H), 1.72-1.67 (m, 2H). LC-MS m/z(ESI) = 408.20 [M+1]
Example 48 -Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)-2-(trifluoromethyl)benzamide (compound 48)
0 HN N N
F C N NO H
Compound 48
NC Step I NC Br Step 2 NC N Step 3 F 3C NH 2 F3 C NH 2 FC N IH 2 48a 48b 48c
0 0
H2N Step 4 H 2N N N FF FC N "'N N)= F3C NH 2 H
48d Compound 48 0 Step 1: 4-Amino-5-bromo-2-(trifluoromethyl)benzonitrile (48b)
4-Amino-2-trifluoromethylbenzonitrile 48a (1.1 g, 5.9 mmol) was dissolved in DMF (10 mL), and NBS (1.05 g, 5.9 mmol) was added. The reaction mixture was stirred at room temperature for 0.5 h and monitored by TLC. The reaction mixture was poured into purified water (100 mL), and a large amounts of solid was precipitated. The solid was collected by filtration and dried under reduced pressure to obtain the title compound 4-amino-5-bromo-2-(trifluoromethyl)benzonitrile 48b (yellow solid, 1.5 g, 96.1% yield). 1HNMR (400 MHz, DMSO-d6) 68.14 (s, 1H), 7.20(s, 1H), 6.90(s, 2H). LC-MS m/z(ESI) = 264.90 [M+1]
Step 2: 4-Amino-5-methyl-2-(trifluoromethyl)benzonitrile (48c)
4-Amino-5-bromo-2-(trifluoromethyl)benzonitrile 48b (1.5 g, 5.66 mmol) was added to a mixture of 1,4-dioxane (20 mL) and water (4 mL), and methylboronic acid (3.4 g, 56.6 mmol), potassium carbonate (2.3 g, 17 mmol) and [1,1' bis(diphenylphosphino)ferrocene]dichloropalladium (417 mg, 0.57 mmol) were added. The system was purged 4 times with nitrogen, and the reaction mixture was refluxed for 5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was purified by column chromatography to obtain the title compound 4-amino-5-methyl-2-(trifluoromethyl)benzonitrile 48c (yellow solid, 850 mg, 75.2% yield). LC-MS m/z(ESI) = 201.20 [M+1]
Step 3: 4-Amino-5-methyl-2-(trifluoromethyl)benzamide (48d)
4-Amino-2-trifluoromethyl-5-methyl-benzonitrile 48c (850 mg, 4.25 mmol) was dissolved in DMSO (2 mL), and potassium carbonate (117 mg, 0.85 mmol) was added, followed by the slow dropwise addition of hydrogen peroxide (3 mL) at room temperature. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was poured into purified water (15 mL), and a large amounts of solid was precipitated. The solid was collected by filtration and dried under reduced pressure to yield the title compound 4-amino-5 methyl-2-(trifluoromethyl)benzamide 48d (yellow solid, 312 mg, 33.6% yield). LC-MS m/z(ESI) = 219.20 [M+1]
Step 4: -Methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)-2-(trifluoromethyl)benzamide (compound 48)
The title compound 5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)-2-(trifluoromethyl)benzamide compound 48 (pale yellow solid, 25 mg, 8.0% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz, DMSO-d6) 6 8.71 (s, 1H), 8.18 (s, 1H), 8.16 (s, 1H),7.83 (s, 1H), 7.44 (s, 1H), 7.38 (s, 1H), 4.44-4.36 (m, 1H), 3.97-3.93 (m, 2H), 3.50-3.35 (m, 2H), 3.32 (s, 3H), 2.47-2.43(m, 2H), 2.36 (s, 3H), 1.75-1.59 (m, 2H). LC-MS m/z (ESI) = 451.20 [M+1]
Example 49 2-Fluoro-4-((9-(3-hydroxy-3-methylcyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 49)
0/ H 2N N N N AJiN N )= F H
Compound 49 OH 0 0 H Step 12HC1 Step 2 H Step 3 N Step 4 N S N NH S~ N NS="N NH
OH OH OH OH OH
Step 5N N Step 6 ) >O Step 7 Step 8 H 2N N N OHH
49f 4g 49h Compound 49
Step 1: Ethyl 4-((trans-3-hydroxycyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate (49b)
trans-3-Aminocyclobutane-1-ol hydrochloride 49a (15.00 g, 303.45 mmol) was dissolved in acetonitrile (200 mL), and potassium carbonate (41.92 g, 303.45 mmol) and ethyl 4-chloro-2-methylthio-5-pyrimidinecarboxylate (28.24 g, 121.38 mmol) were added with stirring at 0 °C. The reaction mixture was warmed to room temperature and stirred for 20 h. The reaction monitored by TLC. After completion of the reaction, a large amount of water was added to the reaction mixture, and a white solid was precipitated. The reaction mixture was filtered through celite, and the filtrate was concentrated to get the title compound ethyl 4-((trans-3-hydroxycyclobutyl)amino)-2 (methylthio)pyrimidine-5-carboxylate 49b (white solid, 30 g, 87.23% yield). H NMR (400 MHz, DMSO-d6) 6 8.54 (s, 1H), 8.29 (d, 1H), 5.14(s, 1H),4.60-4.55 (m,
1H), 4.32-4.25 (m, 3H), 2.47 (s, 3H), 2.24 (t, 4H), 1.30 (t, 3H). LC-MS m/z(ESI)= 284.10 [M+1]
Step 2: 4-((trans-3-Hydroxycyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid (49c)
Ethyl 4-((trans-3-hydroxycyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 49b (30 g, 105.88 mmol) was dissolved in methanol/water (150 mL/150 mL), and sodium hydroxide (6.4 g, 158.82 mmol) was added. The reaction mixture was stirred at room temperature for 12 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate methanol and the pH was adjusted to about 4 with 2 N HCl, and a white solid was precipitated. The reaction mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to obtain the title compound 4-((trans-3 hydroxycyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 49c (white solid, 25 g, 92.49% yield). 'H NMR (400 MHz, DMSO-d6) 6 13.28 (s, 1H), 8.51 (s, 1H), 8.47 (d, 1H), 5.11 (s, 1H), 4.55-4.61 (m,1H), 4.23-4.33 (m, 1H), 2.47 (s, 3H), 2.20-2.26 (m, 4H). LC-MS m/z(ESI) = 256.10 [M+1]
Step 3: 9-(trans-3-Hydroxycyclobutyl)-2-(methylthio)-7,9-dihydro-8H-purin-8-one (49d)
4-((trans-3-Hydroxycyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 49c (25 g, 97.93 mmol) was dissolved in N,N-dimethylacetamide (250 mL), and triethylamine (11.89 g, 117.51 mmol) and diphenylphosphoryl azide (32.34 g, 117.51 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 1 h, heated to 90 °C and refluxed for 5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a large amounts of solid was precipitated. The solid was collected by filtration and dried to obtain the title compound 9-(trans-3-hydroxycyclobutyl)-2-(methylthio)-7,9-dihydro 8H-purin-8-one 49d (white solid, 20 g, 80.95% yield). H NMR (400 MHz, DMSO-d6) 6 11.30 (s, 1H), 8.10 (s, 1H), 5.18 (s, 1H), 5.07-4.99 (m, 1H), 4.56-4.53 (m, 1H), 3.07-3.00 (m, 2H), 2.51(s, 3H), 2.27-2.21 (m, 2H). LC-MS m/z(ESI)= 253.10 [M+1]
Step 4: 9-(trans-3-Hydroxycyclobutyl)-7-methyl-2-(methylthio)-7,9-dihydro-8H-purin-8-one (49e)
9-(trans-3-Hydroxycyclobutyl)-2-(methylthio)-7,9-dihydro-8H-purin-8-one 49d (18 g, 71.35 mmol) was dissolved in N,N-dimethylformamide (300 mL), and dimethyl sulfate (9 g, 71.35 mmol) and cesium carbonate (23.25 g, 71.35 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was slowly added dropwise into ice water with stirring and extracted with ethyl acetate. The organic layer was concentrated and recrystallized from methanol to obtain the title compound 9-(trans-3 hydroxycyclobutyl)-7-methyl-2-(methylthio)-7,9-dihydro-8H-purin-8-one 49e (white solid, 16 g, 84.21% yield).
H NMR (400 MHz, DMSO-d6) 6 8.29 (s, 1H), 5.18 (d, 1H), 5.09-5.01 (m, 1H), 4.56 4.52 (m, 1H), 3.31 (s, 3H), 3.06-2.97 (m, 2H), 2.51 (s, 3H),2.27-2.21 (m, 2H). LC-MS m/z(ESI)= 267.10 [M+1]
Step 5: 7-Methyl-2-(methylthio)-9-(3-oxocyclobutyl)-7,9-dihydro-8H-purin-8-one (49f)
9-(trans-3-Hydroxycyclobutyl)-7-methyl-2-(methylthio)-7,9-dihydro-8H-purin-8-one 49e (2.0 g, 7.51 mmol) was dissolved in dichloromethane (10 mL), Dess-Martin periodinane (4.78 g, 11.26 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was added with saturated sodium bicarbonate and extracted with dichloromethane. The organic layer was concentrated and a solid was precipitated. The reaction mixture was filtered and crystallized to obtain the title compound 7-methyl-2-(methylthio)-9-(3-oxocyclobutyl)-7,9-dihydro-8H purin-8-one 49f (white solid, 1.58 g, 79.6% yield). LC-MS m/z (ESI)= 253.10 [M+1]
Step 6: 9-(3-Hydroxy-3-methylcyclobutyl)-7-methyl-2-(methylthio)-7,9-dihydro-8H-purin-8 one (49g)
7-Methyl-2-(methylthio)-9-(3-oxocyclobutyl)-7,9-dihydro-8H-purin-8-one 49f (1.07 g, 4.05 mmol) was dissolved in tetrahydrofuran (10 mL), and 3 M methylmagnesium bromide (4.7 mL, 14.17 mmol) was slowly added with stirring at 0 °C. The reaction mixture was stirredfor 2.5 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was quenched with saturated aqueous ammonium chloride solution, and extracted with dichloromethane. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 120:1) to obtain the title compound 9-(3-hydroxy 3-methylcyclobutyl)-7-methyl-2-(methylthio)-7,9-dihydro-8H-purin-8-one 49g (white solid, 820 mg, 72.25% yield). 1H NMR (600 MHz, DMSO-d6) 6 8.29 (s, 1H), 5.14 (s, 1H), 4.40-4.32 (m, 1H), 3.31 (s, 3H), 3.02-2.96 (m, 2H), 2.53 (s, 3H), 2.35-2.30 (m, 2H), 1.33 (s, 3H). LC-MS m/z(ESI)= 281.10 [M+1]
Step 7: 9-(3-Hydroxy-3-methylcyclobutyl)-7-methyl-2-(methylsulfinyl)-7,9-dihydro-8H purin-8-one (49h)
9-(3-Hydroxy-3-methylcyclobutyl)-7-methyl-2-(methylthio)-7,9-dihydro-8H-purin-8 one 49g (710 mg, 2.53 mmol) was dissolved in dichloromethane (5 mL), and m chloroperoxybenzoic acid (568 mg, 3.29 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, saturated sodium bicarbonate was added and extracted with dichloromethane. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = :1) to obtain the title compound 9-(3-hydroxy-3-methylcyclobutyl)-7-methyl-2 (methylsulfinyl)-7,9-dihydro-8H-purin-8-one 49h (white solid, 200 mg, 26.65% yield). LC-MS m/z(ESI)= 297.10 [M+1]
Step 8: 2-Fluoro-4-((9-(3-hydroxy-3-methylcyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 49)
9-(3-Hydroxy-3-methylcyclobutyl)-7-methyl-2-(methylsulfinyl)-7,9-dihydro-8H purin-8-one 49h (100 mg, 0.34 mmol) and 4-amino-3-methyl-2-fluorobenzamide intermediate 2 (284 mg, 1.69 mmol) were dissolved in N,N-dimethylformamide (3 mL), and cesium carbonate (330 mg, 1.01 mmol) was added. The reaction mixture was gradually heated to 80 °C and stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into 30 mL of purified water, and extracted with ethyl acetate. After filtrataion, the resultant purified by silica gel column chromatography purification (dichloromethane/methanol (v/v) = 35/1) to obtain the title compound 2-fluoro-4-((9-(3-hydroxy-3-methylcyclobutyl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 49-1 (white solid, 12 mg, 8.88% yield) and compound 49-2 (white solid, 20 mg, 14.80% yield). Compound 49-1: 1H NMR (600 MHz, DMSO-d6) 6 8.60 (s, 1H), 8.21 (s, 1H), 7.87 (d, 1H), 7.55 (d, 1H), 7.46 (s, 1H), 7.42 (s, 1H), 5.13 (s, 1H), 4.41-4.33 (m, 1H), 3.35 (s, 3H), 3.07-2.98 (m, 2H), 2.53 (s, 3H), 2.35-2.30 (m, 2H), 1.36 (s, 3H). LC-MS m/z(ESI)= 401.20 [M+1] Compound 49-2: 1H NMR (600 MHz, DMSO-d6) 6 8.59 (s, 1H), 8.20 (s, 1H), 7.87 (d, 1H), 7.55 (d, 1H), 7.46 (s, 1H), 7.42 (s, 1H), 5.14 (s, 1H), 4.39-4.31 (m, 1H), 3.32 (s, 3H), 3.05-2.97 (m, 2H), 2.54 (s, 3H), 2.33-2.29 (m, 2H), 1.35 (s, 3H). LC-MS m/z(ESI)= 401.20 [M+1]
Example 50 2-Hydroxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 50)
0 HN lN"- N> HO N N N H
Compound 50 0 0 NC H Step I HN NH Step 2 H2N N 10 NH1 2 -" NH 2 HO - N H,
50a 50b 50c
0
Step 3 H 2NHO NN
HO N N N>= H
0 Compound 50 Step 1:
4-Amino-2-ethoxy-5-methylbenzamide (50b)
4-Amino-2-ethoxy-5-methylbenzonitrile 50a (1.0 g, 5.67 mmol) was dissolved in a solvent mixture of ethanol (15 mL) and dimethyl sulfoxide (1 mL), and a 1 N sodium hydroxide solution (3 mL) was added, followed by the slow dropwise addition of hydrogen peroxide (3 mL) at room temperature. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into a saturated sodium sulfite solution (20 mL) and stirred at room temperature for 2 h, and a large amounts of solid was precipitated. The solid was collected by filtration and dried by rotary evaporation under reduced pressure to obtain the title compound 4-amino-2-ethoxy-5-methylbenzamide 50b (yellow solid, 510 mg, 48.6% yield). 'H NMR (400 MHz, DMSO-d6) 6 7.53 (s, 1H), 7.34 (d, 1H), 7.07 (d, 1H), 6.28 (s, 1H), 5.47 (s, 2H), 4.04 (q, 2H), 1.99 (s, 3H), 1.37 (t, 3H). LC-MS m/z(ESI)= 195.20 [M+1]
Step 2: 4-Amino-2-hydroxy-5-methylbenzamide (50c)
4-Amino-2-ethoxy-5-methylbenzamide 50b (510 mg, 2.63 mmol) was dissolved in dichloromethane (2 mL). The reaction mixture was cooled to 0 °C, and boron tribromide solution (13.1 mL, 1 M in THF) was slowly added dropwise. The reaction mixture was warmed to room temperature and stirred for 16 h. The reaction was monitored by TLC. After completion of the reaction,ethyl acetate (20 mL) and saturated ammonium chloride solution (20 mL) were added, and the resulting mixture was stirred for 0.5 h. The organic phase was extracted, concentrated, dried over anhydrous sodium sulfate and filtered, and the filtrate was dried under reduced pressure to obtain the title compound 4-amino-2-hydroxy-5-methylbenzamide 50c (yellow solid, 350 mg, 80.1% yield). 'H NMR (400 MHz, DMSO-d6) 6 12.26 (s, 1H), 7.79 (s, 1H), 7.38 (s, 1H), 7.23 (s, 1H), 5.98 (s, 1H), 5.50 (s, 2H), 1.96 (s, 3H). LC-MS m/z(ESI)= 167.20 [M+1]
Step 3: 2-Hydroxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 50)
The title compound 2-hydroxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran 4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 50 (white solid, 35 mg, 46.9% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz, DMSO-d) 6 12.87 (s, 1H), 8.27 (s, 1H), 8.16-8.13 (m, 2H), 7.64 (s, 1H), 7.61 (s, 1H), 7.54 (s, 1H), 4.47-4.39 (m, 1H), 4.00-3.96 (m, 2H), 3.48-3.37 (m, 2H), 3.33 (s, 3H), 2.57-2.48 (m, 2H), 2.22 (s, 3H), 1.70-1.64 (m, 2H). LC-MS m/z(ESI)= 399.20 [M+1]
Example 51 2-Fluoro-5-methyl-4-((7-methyl-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)methylbenzoate (compound 51)
O N N H b Compound 51
0 0 0I
O N N O Step 1 NH2 Step 2 HO F N NH, O F NH, N' NN Fa N H
36a 51a Compound 51 0
Step 1: Methyl 4-amino-2-fluoro-5-methylbenzoate (51a)
4-Amino-2-fluoro-5-methylbenzoic acid 36a (1.0 g, 5.91 mmol) was dissolved in methanol (6 mL), and 4 drops of concentrated sulfuric acid were added. The reaction mixture was heated to 60 °C and refluxed for 2 h. The reaction was monitored by TLC. After completion of the reaction, saturated sodium bicarbonate solution (20 mL) were added and extracted with ethyl acetate, and the organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate and concentrated, and the residue was purified by column chromatography (PE:EA (v/v) = 5:1) to obtain the title compound methyl 4-amino-2-fluoro-5-methylbenzoate 51a (pale yellow solid, 0.8 g, 43.7% yield). LC-MS m/z(ESI) = 184.20 [M+1]
Step 2: 2-Fluoro-5-methyl-4-((7-methyl-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)methylbenzoate (compound 51)
The title compound 2-fluoro-5-methyl-4-((7-methyl-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)methylbenzoate compound 51 (white solid, 239 mg, 79.8% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz, DMSO-d) 8.62 (s, 1H), 8.24 (s, 1H), 8.07 (d, 1H), 7.69 (d, 1H), 4.49-4.41 (m, 1H), 4.00-3.96 (m, 2H), 3.81 (s, 3H), 3.49-3.36 (m, 2H), 3.34 (s, 3H), 3.17 (s, 3H), 2.65-2.50 (m, 2H), 1.74-1.65 (m, 2H). LC-MS m/z(ESI) = 402.20 [M+1]
Example 52 2-Fluoro-5-methyl-4-((7-methyl-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)benzoic acid (compound 52)
0 HO N NN FN NN N H
Compound 52 d
0 0 ON
0 N HO -O - - F NN H N>= FaN N>= N H
b 0 Compound 51 Compound 52
2-Fluoro-5-methyl-4-((7-methyl-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)methylbenzoate compound 51 (300 mg, 0.75 mmol) was dissolved in tetrahydrofuran (3 mL), and a sodium hydroxide solution (80 mg in 3 mL of water) was added. The reaction mixture was heated to 50 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the pH of reaction mixture was adjusted to about 5 with 2 N hydrochloric acid and concentrated under reduced pressure to remove tetrahydrofuran, and a large amounts of solid was precipitated. The solid was collected by filtration and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to obtain the title compound 2-fluoro-5 methyl-4-((7-methyl-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzoic acid compound 52 (white solid, 162 mg, 56.0% yield). 'H NMR (400 MHz, DMSO-d6) 69.14 (s, 1H), 8.29 (s, 1H), 7.94 (d, 1H), 7.71 (d, 1H), 4.49-4.41 (m, 1H), 4.00-3.96 (m, 2H), 3.46-3.39 (m, 2H), 3.34 (s, 3H), 2.67-2.43 (m, 2H), 2.33 (s, 3H), 1.76-1.67 (m, 2H). LC-MS m/z(ESI) = 388.20 [M+1]
Example 53 2-Ethoxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 53)
0 HN N O "-'O hNN N H
Compound 53 0 HN N HNNN=0 H 2N *C N - H N N O H tO NH2 H 0 0 Intermediate 1 Compound 53 50b
The title compound 2-ethoxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4 yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 53 (pale yellow solid, 35 mg, 12.1% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz, DMSO-d) 68.38 (s, 1H), 8.18 (s, 1H), 7.76 (s, 1H), 7.71 (s, 1H), 7.51 (d, 1H), 7.41 (d, 1H), 4.48-4.40 (m, 1H), 4.17 (q, 2H), 3.98-3.95 (m, 2H), 3.47 3.36 (m, 2H), 3.34 (s, 3H), 2.57-2.53 (m, 2H), 2.24 (s, 3H), 1.72-1.63 (m, 2H), 1.41 (t, 3H). LC-MS m/z(ESI) = 413.20 [M+1]
Example 54 N,4-Dimethyl-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)picolinamide (compound 54)
N O H b Compound 54
0 NN N0
N CI N + N H N NH N N' N)= O NH 2 H
0 0 54a Intermediate I Compound 54 The title compound N,4-dimethyl-5-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl) 8,9-dihydro-7H-purin-2-yl)amino)picolinamide compound 54 (pale yellow solid, 30 mg, 23.2% yield) was prepared according to the synthesis method of compound 1. 'H NMR (400 MHz DMSO) 69.84 (s, 1H), 8.93 (d, 2H), 8.26 (d, 1H), 8.13 (s, 1H), 4.44-4.38 (m, 1H), 3.97-3.93 (m, 2H), 3.44-3.38 (m, 2H), 3.34 (s, 3H), 2.83 (d, 3H), 2.43-2.37 (m, 5H), 2.15 (s, 3H), 1.68 (dd, 2H). LC-MS m/z(ESI) = 398.10 [M+1]
Example 55 7-Methyl-2-((4-methyl-6-oxo-1,6-dihydropyridin-3-yl)amino)-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 55)
H N N N H
0 Compound 55
0 Step 0 Step2 0 N 2LN HNep >==O HN NO NHH N N N 02 2 H
55a 55b Compound 55 0 Step 1: -Amino-4-methylpyridin-2(1H)-one (55b)
4-Methyl-5-nitropyridine-2(1H)-one 55a (5.0 g, 32.44 mmol) and iron powder (9.0 g, 162.2 mmol) were added to a dry reaction flask and well mixed with ethanol/water=10/1 (110 mL), and then hydrochloric acid (7 mL, 2 mol/L) was added dropwise. The reaction mixture was stirred at 85 °C for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filter cake was washed with an appropriate amount of ethanol. The filtrate was concentrated, the pH was adjusted to about 8 with saturated sodium bicarbonate and purified by silica gel column chromatography (dichloromethane:methanol (v/v)= 10:1) to give the title compound 5-amino-4-methylpyridin-2(1H)-one 55b (dark green solid, 2.5 g, 62.08% yield). 1H NMR (400 MHz, DMSO-d6) 6 10.63 (s, 1H), 6.75 (s, 1H), 6.11 (s, 1H), 4.06 (s, 2H), 2.02 (s, 3H).
LC-MS m/z (ESI) = 125.10 [M+1]
Step 2: 7-Methyl-2-((4-methyl-6-oxo-1,6-dihydropyridin-3-yl)amino)-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 55)
2-Chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 55b (200 mg, 0.74 mmol), 5-amino-4-methylpyridin-2(1H)-one (185 mg, 1.48 mmol), cesium carbonate (525 mg, 1.48 mmol) and Brettphos G3 Pd (67 mg, 0.074 mmol) were added to a dry reaction flask, followed by the addition of 1,4-dioxane (20 mL). The system was purged three times with nitrogen, and the reaction mixture was stirred at 110 °C for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 20/1) to obtain the title compound 7-methyl-2-((4-methyl-6-oxo-1,6-dihydropyridin-3-yl)amino)-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 55 (yellow solid, 38 mg, 10.02% yield). 1H NMR (400 MHz, DMSO-d) 6 11.29 (s, 1H), 8.19 (s, 1H), 7.96 (s, 1H), 7.25 (s, 1H), 6.21 (s, 1H), 4.40-4.32 (m, 1H), 3.97-3.93 (m, 2H), 3.44-3.36 (m, 2H), 3.26 (s, 3H), 2.47-2.42 (m, 2H), 2.01 (s, 3H), 1.64-1.60 (m, 2H) LC-MS m/z (ESI) = 357.20 [M+1]
Example 56 2-((1,4-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 56)
O' N N ANN 'j NN N H
Compound 56
H Step Itep 2Step S3 2 NI O H N02 A N ~~N- N NH 2 N H N N 2
55a 56a 56b Compound 56
Step 1: 1,4-Dimethyl-5-nitro-3,4-dihydropyridin-2-2(1H)-one (56a)
4-Methyl-5-nitropyridine-2(1H)-one 55a (5.0 g, 32.44 mmol) and potassium carbonate (9.0 g, 48.66 mmol) were added to a dry reaction flask and well mixed with DMF, and iodomethane (3 mL, 64.88 mmol) was added dropwise at 0 °C. The reaction mixture was warmed to room temperature and stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, the filtrate was extracted with water and ethyl acetate, and the organic phases were combined, dried and concentrated to obtain the title compound 1,4-dimethyl-5-nitro-3,4-dihydropyridin 2-2(1H)-one 56a (yellow solid, 5.2 g, 95.41% yield).
'HNMR(400 MHz, Chloroform-d)6 8.62 (s, 1H), 6.39 (s, 1H), 3.63 (s, 3H),2.56-2.53 (m, 3H). LC-MS m/z (ESI) = 169.10 [M+1]
Step 2: -Amino-1,4-dimethylpyridin-2(1H)-one (56b)
1,4-Dimethyl-5-nitropyridine-2(1IH)-one 56a (5.2 g, 30.92 mmol) and iron powder (9.0 g, 154.62 mmol) were added to a dry reaction flask and well mixed with ethanol/water (v/v=10/1, 110 mL), and then hydrochloric acid (10 mL, 2 mol/L) was added. The reaction mixture was stirred at 85 °C for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filter cake was washed with an appropriate amount of ethanol. The filtrate was concentrated, the pH was adjusted to about 8 with saturated sodium bicarbonate, and purified by silica gel column chromatography (dichloromethane:methanol (v/v) = 10:1) to give the title compound 5-amino-1,4-dimethylpyridin-2(1H)-one 56b (dark green solid, 2.5 g, 58.55% yield). 1H NMR (400 MHz, DMSO-d6) 6 6.88 (s, 1H), 6.16 (s, 1H), 3.81 (s, 2H), 3.30 (s, 3H), 2.03 (s, 3H). LC-MS m/z (ESI) = 141.10 [M+1]
Step 3: 2-((1,4-Dimethyl-6-oxo-1,6-dihydropyridin-3-yl)amino)-7-methyl-9-(tetrahydro-2H pyran-4-yl)-7,9-dihydro-8H-purin-8-one (compound 56)
2-Chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 56b (200 mg, 0.74 mmol), 5-amino-1,4-dimethyl-3,4-dihydropyridin-2(1H)-one (206 mg, 1.48 mmol), cesium carbonate (525 mg, 1.48 mmol) and Brettphos G3 Pd (67 mg, 0.074 mmol) were added to a dry reaction flask, followed by the addition of 1,4-dioxane (20 mL). The system was purged three times with nitrogen, and the reaction mixture was stirred at 110 °C for 2.5 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1) to obtain the title compound 7-methyl-2-((4-methyl-6-oxo-1,6-dihydropyridin-3-yl)amino)-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one compound 56 (yellow solid, 97 mg, 35.36% yield). 1H NMR (400 MHz, DMSO-d) 68.26 (s, 1H), 7.98 (s, 1H), 7.66 (s, 1H), 6.27 (s, 1H), 4.41-4.33 (m, 1H), 3.98-3.94 (m, 2H), 3.42 (d, 2H), 3.38 (s, 3H), 3.27 (s, 3H), 2.54 2.44 (m, 2H), 2.01 (s, 3H), 1.65-1.62 (m, 2H). LC-MS m/z (ESI) = 371.20 [M+1]
Example 57 2-Methoxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 57)
0 H 2N N~' H N O H
d Compound 57
N Step 1 Step 2 H2NH F NH 2 0 NYH 2 No NH2
2A 57a 57b
0
Step 3 H 2N O N0 a'N NN N) - O H
Compound 57 Step 1: 4-Amino-2-methoxy-5-methylbenzonitrile (57a)
4-Amino-2-fluoro-5-methylbenzonitrile 2A (1.0 g, 6.67 mmol) was dissolved in a mixture of methanol (10 mL) and purified water (2 mL), and then potassium hydroxide (1.50 g, 26.68 mmol) was added. The reaction mixture was refluxed for 4 h. The reaction was monitored by TLC. After completion of the reaction, 20 mL of purified water was added, and the resulting mixture was concentrated under reduced pressure to remove methanol and then extracted 3 times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate and concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate (v/v) = 5:1) to obtain 4-amino-2-methoxy-5-methylbenzonitrile 57a (white solid, 300 mg, 27.8% yield). LC-MS m/z(ESI) = 163.10 [M+1]
Step 2: 4-Amino-2-methoxy-5-methylbenzamide (57b)
4-Amino-2-methoxy-5-methylbenzonitrile 57a (300 mg, 1.85 mmol) and potassium carbonate (51 mg, 0.37 mmol) were dissolved in DMSO (1 mL), and the mixture was added slowly dropwise with hydrogen peroxide (2 mL) at room temperature, and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into purified water (10 mL), and a large amounts of solid was precipitated. The solid was collected by filtration and dried under reduced pressure to obtain the title compound 4-amino-2-methoxy-5-methylbenzamide 57b (yellow solid, 256 mg, 76.7% yield). LC-MS m/z(ESI) = 181.10 [M+1]
Step 3: 2-Methoxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 57)
The title compound 2-methoxy-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran 4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 57 (white solid, 35 mg, 12.1% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz, DMSO-d6) 68.39 (s, 1H), 8.19 (s, 1H), 7.78 (s, 1H), 7.70 (s, 1H), 7.55 (s, 1H), 7.37 (s, 1H), 4.47-4.41 (m, 1H), 3.98-3.94(m, 2H), 3.91(s, 3H), 3.44-3.38 (m, 2H), 3.33 (s, 3H), 2.60-2.52 (m, 2H), 2.25 (s,3H), 1.68 (d, 2H). LC-MS m/z(ESI) = 413.20 [M+1]
Example 58 -Methyl-6-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)nicotinamide (compound 58)
0
H2 N N N N N<IN' N>= H
Compound 58
0 0 NC NH2 NCStepi1 H 2N j Step 2 H 2N N >0 "IN N N N>= N NH 2 N- NH 2 H b 0 20a 58a Compound 58 Step 1: 6-Amino-5-methylnicotinamide (58a)
6-Amino-5-methylnicotinonitrile 20a (300 mg, 2.2 mmol) and potassium carbonate (46 mg, 0.33 mmol) were dissolved in 1 mL of dimethyl sulfoxide, and 300 pL of hydrogen peroxide was added in an ice bath. Then the reaction mixture was gradually heated to °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, 5 mL of water was added to the reaction mixture, and a white solid was precipitated. The mixture was filtered, and the filter cake was concentrated to dryness under reduced pressure to obtain the title compound 6-amino-5-methylnicotinamide 58a (white solid, 200 mg, 58% yield). 1H NMR (400 MHz DMSO) 68.34 (s, 1H), 7.68 (s, 1H), 7.63 (s, 1H), 6.98 (s, 1H), 6.25
(s, 1H), 2.04 (s, 3H). LC-MS m/z(ESI)= 152.10 [M+1]
Step 2: -Methyl-6-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)nicotinamide (compound 58)
The title compound 5-methyl-6-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)nicotinamide compound 58 (white solid, 17 mg, 13.1% yield) was prepared according to the synthesis method of compound 1. 1H NMR (400 MHz DMSO) 69.19 (s, 1H), 8.59 (s, 1H), 8.12 (s, 1H), 8.00 (s, 1H), 7.96
(s, 1H), 7.38 (s, 1H), 4.45-4.35 (m, 1H), 3.94 (dd, 2H), 3.40-3.30 (m, 2H), 3.33 (s, 3H), 2.50 (s, 2H), 2.22(s,3H), 1.63(dd, 2H). LC-MS m/z(ESI)= 384.20 [M+1]
Example 59 -Chloro-2-fluoro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 59)
H 2N CI N F N<N N>== H
Compound 59
0 0 NC N2 Step1 NC CI Step 2 H 2N C Step 3 H 2N -N N N N
b -~F
F NH, F NH 2 F NH 2 H 59a 59b 59c Compound 59 0
Step 1: 4-Amino-5-chloro-2-fluorobenzonitrile (59b)
4-Amino-2-fluorobenzonitrile 59a (10.0 g, 73.46 mmol) was dissolved in acetonitrile (100 mL), and N-chlorosuccinimide (10.79 g, 80.81 mmol) was added in an ice bath. The reaction mixture was gradually warmed to room temperature, heated to 80 °C and stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured directly into 400 mL of water and extracted with 200 mL of ethyl acetate; the organic layer was collected, dried and concentrated to obtain the title compound 4-amino-5-chloro-2-fluorobenzonitrile 59b (white solid, 6 g, 46.7% yield). 1HNMR (400 MHz, DMSO-d6) 6 7.79-7.77 (m, 2H), 6.78(s, 2H), 6.66 (d, 1H). LC-MS m/z(ESI)= 171.10 [M+1]
Step 2: 4-Amino-5-chloro-2-fluorobenzamide (59c)
4-Amino-5-chloro-2-fluorobenzonitrile 59b (500 mg, 2.93 mmol) was dissolved in 1 M sodium hydroxide (14.66 mL) and methanol (1.19 mL). The solution was heated to °C, and then 30% hydrogen peroxide (0.3 mL) was added dropwise. The reaction mixture was reacted for 16 h with the temperature maintained. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was directly diluted with water (50 mL) and extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried and concentrated, and the residue was purified by silica gel column chromatography to obtain the title compound 4 amino-5-chloro-2-fluorobenzamide 59c (280 mg, white solid, 50.7% yield). 1H NMR (400 MHz, CDC3-d6) 67.57 (d, 2H), 7.36 (s, 1H), 7.20 (s, 1H), 6.53 (d, 2H), 6.18 (s, 2H). LC-MS m/z(ESI)= 189.10 [M+1] Step 3: -Chloro-2-fluoro-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 59)
4-Amino-5-chloro-2-fluorobenzamide 59c (280.74 mg, 1.49 mmol), 2-chloro-7 methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one intermediate 1 (200 mg, 0.74 mmol), cesium carbonate (485 mg, 1.49 mmol), and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (67.4 mg, 0.07 mmol) were dissolved in dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at
100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to yield compound 5-chloro-2-fluoro-4-((7-methyl-8-oxo-9-(tetrahydro 2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 59 (white solid, 40 mg, 12.77 % yield). 1H NMR (401 MHz, DMSO-d6) 6 8.44 (s, 1H), 8.29 (d, 1H), 8.25 (s, 1H), 7.77 (d, 1H), 7.64 (s, 1H), 7.60 (s, 1H), 4.49-4.43 (m, 1H), 4.00-3.97 (m, 2H), 3.48-3.44 (m, 2H), 3.34 (s, 3H), 2.57-2.53 (m, 2H), 1.72-1.68 (m, 2H). LC-MS m/z(ESI)= 421.10 [M+1]
Example 60 2-Fluoro-3,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 60)
0 H2N - N NO F N 'N N H
Compound 60
NC Step 1 NC Cl Step 2 NC N Step 3
F NH2 F NH 2 F NH 2 CI
59a 60a 60b
N0 NC N ON Step 4 H2 N N O H HNH bN
0 0 60c Compound 60
Step 1: 4-Amino-3,5-dichloro-2-fluorobenzonitrile (60a)
4-Amino-2-fluorobenzonitrile 59a (10.0 g, 73.46 mmol) was dissolved in acetonitrile (100 mL), and N-chlorosuccinimide (21 g, 160 mmol) was added in an ice bath. The reaction mixture was gradually warmed to room temperature, heated to 80 °C and reacted for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured directly into 400 mL of water and extracted with 200 mL of ethyl acetate; the organic layer was collected, dried and concentrated to afford the title compound 4-amino-3,5-chloro-2-fluorobenzonitrile 60a (white solid, 7 g, 46.5% yield). 1H NMR (400 MHz, DMSO-d6) 6 7.86 (d, 1H), 7.03 (s, 2H).
LC-MS m/z(ESI)= 205.10 [M+1] Step 2: 4-Amino-2-fluoro-3,5-dimethylbenzonitrile (60b)
4-Amino-3,5-dichloro-2-fluorobenzonitrile 60a (7 g, 34.14 mmol), methylboronic acid
(20.44 g, 341.44 mmol), potassium phosphate (36.24 g, 170.72 mmol), 2 dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (4.88 g, 10.24 mmol), and palladium(II) acetate (1.15 g, 5.12 mmol) were dissolved in dioxane (100 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography to obtain the title compound 4-amino-2-fluoro-3,5 dimethylbenzonitrile 60b (white solid, crude product, 6 g, 107% yield). 'H NMR (400 MHz, CDCl3-d6) 6 7.14 (d, 1H), 5.94 (s, 2H), 1.99 (s, 3H), 1.94 (d, 2H). LC-MS m/z(ESI)= 165.10 [M+1]
Step 3: 2-Fluoro-3,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzonitrile (60c)
4-Amino-2-fluoro-3,5-dimethylbenzonitrile 60b (1.22 g, 7.44 mmol), 2-chloro-7 methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one intermediate 1 (2.0 g, 7.44 mmol), cesium carbonate (4.85 g, 14.89 mmol), and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (674 mg, 0.74 mmol) were dissolved in dioxane (20 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography to obtain compound 2-fluoro-3,5-dimethyl-4-((7 methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzonitrile 60c (413 mg, yellow solid, 14 % yield). H NMR (401 MHz, DMSO-d6) 6 9.02 (s, 1H), 8.12 (s, 1H), 7.91 (d, 1H), 4.49-4.39 (m, 1H), 4.00-3.96 (m, 2H), 3.45-3.39 (m, 2H), 2.32 (s, 3H), 2.22 (d, 3H), 1.71-1.65 (m, 2H). LC-MS m/z(ESI)= 397.10 [M+1]
Step 4: 2-Fluoro-3,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide(compound 60)
2-Fluoro-3,5-dimethyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzonitrile 60c (413 mg, 1.04 mmol) was dissolved in 1 M sodium hydroxide (5 mL) and methanol (10 mL). The solution was heated to 60 °C, and then 30% hydrogen peroxide (0.16 mL) was added dropwise. The reaction mixture was stirred for 4 h with the temperature maintained. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was directly diluted with water (50 mL) and extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried and concentrated, and the residue was purified by silica gel column chromatography to obtain the title compound 2-fluoro-3,5-dimethyl 4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide compound 60 (white solid, 40 mg, 9.3% yield). 'H NMR (400 MHz, CDC3-d6) 6 8.65 (s, 1H), 7.91 (s, 1H), 7.60 (s, 1H), 7.54 (s, 1H), 7.36 (d, 1H), 4.42-4.34 (m, 1H), 3.98-3.94 (m, 1H), 3.43-3.37 (m, 2H), 3.26 (s, 3H), 2.13 (s, 3H), 2.04 (d, 3H), 1.67-1.63 (m, 2H). LC-MS m/z(ESI)= 415.10 [M+1]
Example 61 4-((9-(trans-3-Cyanocyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 2-fluoro-5-methylbenzamide (compound 61)
0/ H2 N N N
H
bN Compound 61 0 0 0 0 H
N - " IN"I'l OH So4 IN" IN Step 1~ N - - Step4 N N H Step S Step 2 Step 3 N NHISN NH INNH
COOH CONH 2 CN CNN N
1150 61a 61b 61c 61d
Step O Step 6 l O, Step 7 H2N N NN> - - -'SN>N.j) INNN FN
CN CN CtN 61f 61g Compound 61 Step 1: Ethyl 4-((trans-3-carbamoylcyclobutyl)amino)-2-(methylthio)pyrimidine-5 carboxylate (61b)
The starting material trans-3-((5-(ethoxycarbonyl)-2-(methylthio)pyrimidin-4 yl)amino)cyclobutane-1-carboxylic acid 61a (13.0 g, 41.8 mmol) was added to dichloromethane (130 mL), andNN,N,NN-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (19.1 g, 50.2 mmol) and triethylamine (16.9 g, 167.2 mmol) were added. The mixture was stirred at room temperature for 10 min, and ammonium chloride (6.7 g, 125.4 mmol) was added. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, 50 mL of saturated aqueous sodium bicarbonate and dichloromethane was added to the reaction mixture. The organic layer was concentrated over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude title compound ethyl 4-((trans-3 carbamoylcyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 61b (yellow viscous liquid, 12.5 g, 96.5% yield). LCMS m/z(ESI) = 311.20 [M+1]
Step 2: Ethyl 4-((trans-3-cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate (61c)
Ethyl 4-((cis-3-carbamoylcyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 61b (12.5 g, 4.03 mmol) was added to dichloromethane (130 mL), and pyridine (13.2 g, 167.2 mmol) and trifluoroacetic anhydride (26.3 g, 125.4 mmol) were added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the pH of reaction mixture was adjusted to 3 with 2 N hydrochloric acid and extracted with dichloromethane. Dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was then purified by column chromatography (petroleum ether/ethyl acetate (v/v) = 5:1) to give the title compound ethyl 4-((trans-3-cyanocyclobutyl)amino)-2 (methylthio)pyrimidine-5-carboxylate 61c (white solid, 1.7 g, 10.2% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.65 (s, 1H), 8.46 (d, 1H), 4.70-4.61 (m, 1H), 4.37 4.31 (m, 2H), 2.92-2.81 (m, 3H), 2.52 (s, 3H), 2.46-2.35 (m, 2H), 1.38(t, 3H). LC-MS m/z(ESI)= 293.20 [M+1]
Step 3: 4-((trans-3-Cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid (61d)
Ethyl 4-((cis-3-Cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 61c (1.7 g, 5.8 mmol) was dissolved in a mixture of tetrahydrofuran (10 mL) and water (10 mL), and lithium hydroxide (487 mg, 11.6 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 4-((trans-3-cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 61d (white solid, 1.5 g, 97.4% yield). LC-MS m/z(ESI)= 265.10 [M+1]
Step 4: trans-3-(2-(Methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile (61e)
4-((trans-3-Cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 61d (1.5 g, 5.7 mmol) was dissolved in N,N-dimethylformamide (15 mL), and triethylamine (576 mg, 5.7 mmol) and diphenylphosphoryl azide (1.57 g, 5.7 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 5:1 to 1:10) to obtain the title compound trans-3 (2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 61e (white solid, 1.0 g, 67% yield). 1H NMR (400 MHz, DMSO-d6) 6 11.35 (s, 1H), 8.11 (s, 1H), 4.92-3.92 (m, 1H), 3.37 3.26 (m, 2H), 3.27-3.15 (m, 1H), 2.78-2.67 (m, 2H), 2.58 (s, 3H). LC-MS m/z(ESI)= 262.10 [M+1]
Step 5: trans-3-(7-Methyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (61f)
trans-3-(2-(Methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 61e (500 mg, 19.1 mmol) was dissolved in dimethylformamide (5 mL), and dimethyl sulfate (2.4 g, 19.1 mmol) and cesium carbonate (12.5 g, 38.2 mmol) were added at °C. The reaction mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, then 50 mL of water was added, and a solid was precipitated. The solid was collected by filtration and dried by rotary evaporation under reduced pressure to obtain the title compound the title compound trans-3-(7-methyl-2 (methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 61f (white solid, 462 mg, 87.7% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.61 (s, 1H), 4.90-4.02 (m, 1H), 3.34(s, 3H), 3.37 3.25(m, 2H), 3.25-3.12(m, 1H), 2.89(s, 3H), 2.86-2.75(m, 2H). LC-MS m/z (ESI)= 276.10 [M+1]
Step 6: trans-3-(7-Methyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane 1-carbonitrile (61g)
trans-3-(7-Methyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 61f (462 mg, 1.67 mmol) was dissolved in methanol (10 mL), and 2 mL of purified water was added, followed by the addition of potassium peroxymonosulfate (1.02 g, 1.67 mmol). The reaction mixture was heated to 60 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove methanol, and 5 mL of purified water was added. The resulting mixture was extracted with dichloromethane (20 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation under reduced pressure to remove the solvent to obtain the title compound trans-3-(7-methyl-2 (methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 61g (pale yellow solid, 181 mg, 35.9% yield). LC-MS m/z(ESI)= 308.10 [M+1]
Step 7: 4-((9-(trans-3-Cyanocyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 2-fluoro-5-methylbenzamide (compound 61)
trans-3-(7-Methyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane 1-carbonitrile 61g (181 mg, 0.59 mmol) and 4-amino-3-methyl-2-fluorobenzamide (538 mg, 3.20 mmol) were dissolved in N,N-dimethylformamide (3 mL), followed by nitrogen purging and the addition of potassium tert-butoxide (143 mg, 1.28 mmol). The reaction mixture was stirred under nitrogen at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was poured into 30 mL of purified water, and a large amounts of yellow solid was precipitated. The solid was collected by filtration and purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(trans-3-cyanocyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 2-fluoro-5-methylbenzamide compound 61 (yellow solid, 27 mg, 11.6 % yield). 'H NMR (400 MHz, DMSO-d6) 6 8.62(s, 1H), 8.20(s, 1H), 7.88(d, 1H), 7.55(d, 1H), 7.49(s, 1H), 7.40(s, 1H), 5.14-5.01(m, 1H), 3.64-3.59(m, 1H), 3.31(s, 3H)3.26-3.21(m, 2H), 2.78-2.70(m, 2H), 2.32(s, 3H). LC-MS m/z(ESI)= 396.20 [M+1]
Example 62 4-((7-Ethyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 62)
0
H 2N N N
F N> N N H
Compound 62
H O HH
1D 62a Compound 62 Step 1: 2-Chloro-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro- 8H-purin- 8-one (62a)
2-Chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 -one 1D (400 mg, 1.57 mmol) was dissolved in NN-dimethylformamide (8 mL), and cesium carbonate (511 mg, 1.57 mmol) and iodoethane (293 mg, 1.88 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction,then 10 mL of water was added, and the resulting mixture was extracted 3 times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated. The reaction mixture was concentrated by rotary evaporation under reduced pressure to remove organic solvent to obtain the title compound 2-chloro-7 ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro- 8H-purin- 8-one 62a (white solid, 290 mg, 65.32% yield). 1 H NMR (400 MHz, DMSO-d6) S 8.44 (s, 1H), 4.50-4.41 (m, 1H), 3.99-3.95 (m, 2H), 3.89 (q, 2H), 3.45 (t, 2H), 2.46-2.41 (m, 2H), 1.71-1.67 (m, 2H), 1.25 (t, 3H). LC-MS m/z(ESI)= 283.10 [M+1]
Step 2: 4-((7-Ethyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 62)
2-Chloro-7-ethyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 62a (150 mg, 0.53 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (350 mg, 2.12 mmol), cesium carbonate (690 mg, 2.12 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl- 1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (72 mg, 0.08 mmol) were dissolved in 1,4-dioxane (5 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1), and purified by Pre-HPLC to obtain the title compound 4-((7-ethyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 62 (white solid, 38 mg, 17.28% yield). 1H NMR (400 MHz, DMSO-d6) 68.53 (s, 1H), 8.25 (s, 1H), 7.89 (d, 1H), 7.55 (d, 1H), 7.42 (d, 2H), 4.48-4.40 (m, 1H), 3.98 (dd, 2H), 3.85 (q, 2H), 3.43 (t, 2H), 2.58-2.54 (m, 2H), 2.30 (s, 3H) 1.71-1.68 (m, 2H), 1.25 (t, 3H). LC-MS m/z(ESI) = 415.20 [M+1]
Example 63 2-Fluoro-4-((7-isopropyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)-5-methylbenzamide (compound 63)
0 H2N N N -=O 1 F N ',N N H
Compound 63
H0 N N NN0 C N Step I O Step 2 H 2N N NO= CI a'N>== CI N NNCN K-C NNN H
0 0 0 1D 63a Compound 63 Step 1: 2-Chloro-7-isopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (46a)
2-Chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 1D (520 mg, 2.04 mmol) was dissolved in N,N-dimethylformamide (10 mL), and cesium carbonate (665 mg, 2.04 mmol) and 2-iodopropane (416 mg, 2.45 mmol) were added at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction,then 20 mL of water was added, and the resulting mixture was extracted 3 times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated to afford the title compound 2-chloro-7-isopropyl-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 63a (white solid, 465 mg, 76.74% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.52 (s, 1H), 4.65-4.56 (m, 1H), 4.49-4.40 (m, 1H), 3.99-3.95 (m, 2H), 3.44 (t, 2H), 2.46-2.42 (m, 2H), 1.70-1.67 (m, 2H), 1.43 (d, 6H). LC-MS m/z(ESI) = 297.10 [M+1]
Step 2: 2-Fluoro-4-((7-isopropyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)-5-methylbenzamide (compound 63)
The title compound 2-fluoro-4-((7-isopropyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 63 (white solid, 58 mg, 20.06% yield) was prepared according to the synthesis method of compound 62. 1H NMR (400 MHz, Chloroform-d) 6 10.17 (s, 1H), 7.98 (d, 1H), 7.93 (s, 1H), 7.76 (d, 1H), 6.79 (d, 1H), 6.39 (s, 1H), 4.73 (t, 1H), 4.52 (t, 1H), 4.11 (dd, 2H), 3.49 (t, 2H), 2.63-2.55 (m, 1H), 2.40 (s, 3H), 1.75 (d, 2H), 1.51 (d, 6H). LC-MS m/z(ESI) = 429.20 [M+1]
Example 64 4-((7-Cyclopropyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide(compound64)
0 H2 N N NO F NAN N H
Compound 64
H 0 N N N O>= Step 2 H2N N N Step 1 I CI CN N O=0 ~CI N N F JNN N H 00b 0 1D 64a Compound 64 Step 1: 2-Chloro-7-cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (64a)
2-Chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purine-8-one ID (1 g, 3.9 mmol), cyclopropylboronic acid (676 mg, 7.8 mmol), anhydrous copper(r) acetate (715 mg, 3.9 mmol), potassium carbonate (1.06 g, 7.8 mmol) and 1,10-phenanthroline (710 mg, 3.9 mmol) were dissolved in 1,2-dichloroethane (12 mL), and the reaction mixture was stirred open to air, then heated to 70 °C and stirred for 6 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 3:1) to obtain the title compound 2-chloro-7 cyclopropyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 64a (white solid, 690 mg, 56 % yield). 1H NMR (400 MHz CDCI3) 6 8.15 (s, 1H), 4.59-4.51 (m, 1H), 4.12 (dd, 2H), 3.55-3.49 (m, 2H), 2.96-2.91 (m, 1H), 2.77-2.67 (m, 2H), 1.77-1.68 (m, 2H), 1.26-1.13 (m, 2H), 1.04-1.00(m, 2H). LC-MS m/z(ESI) = 295.20 [M+1]
Step 2: 4-((7-Cyclopropyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 64)
The title compound 4-((7-cyclopropyl-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 64 (white solid, 20 mg, 16.2% yield) was prepared according to the synthesis method of compound 62. 1H NMR (400 MHz DMSO) 6 8.56 (s, 1H), 8.11 (s, 1H), 7.88 (d, 1H), 7.54 (d, 1H), 27.43 (d, 2H), 5.98-5.87 (m, 1H), 5.24-5.19 (m, 2H), 4.49-4.42 (m, 3H), 3.99 (dd, 2H), 3.43 (t, 2H), 2.59-2.51 (m, 2H), 2.29 (s, 1H), 1.71 (dd, 2H). LC-MS m/z(ESI) = 427.20 [M+1]
Example 65 4-((9-(trans-3-Cyanocyclobutyl)-7-ethyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 65)
0 H2 N N NO FaNAN N
bN Compound 65 H 0 N N N
Stp S Step N i. >=0 - Stop2 S N N - Step3 H2N ~ N1:N F NN
CN SNC bN CN -N 61e 65a 65b Compound 65 Step 1: trans-3-(7-Ethyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (65a)
trans-3-(2-(Methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 61e (500 mg, 19.1 mmol) was dissolved in dimethylformamide (5 mL), and iodoethane (3.0 g, 19.1 mmol) and sodium hydroxide (1.53 g, 38.2 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, 50 mL of water was added, and a solid was precipitated. The reaction mixture was filtered and concentrated by rotary evaporation under reduced pressure to obtain the title compound trans-3-(7-ethyl-2-(methylthio)-8 oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 65a (white solid, 457 mg, 82.5% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.68 (s, 1H), 4.97-4.87 (m, 1H), 3.93(q, 2H), 3.34(s, 3H), 3.33-3.23(m, 2H), 2.91(s, 3H), 2.74-2.59(m, 2H), 1.27(t, 3H). LC-MS m/z (ESI)= 290.10 [M+1]
Step 2: trans-3-(7-Ethyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (65b)
Trans-3-(7-Ethyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 65a (457 mg, 1.58 mmol) was dissolved in methanol (10 mL), and 2 mL of purified water was added, followed by the addition of potassium peroxymonosulfate (1.00 g, 1.58 mmol). The reaction mixture was heated to 60 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove methanol, and 5 mL of purified water was added. The resulting mixture was extracted 3 times with dichloromethane (20 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated by rotary evaporation under reduced pressure to remove the solvent and thus to obtain trans-3-(7-ethyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H purin-9-yl)cyclobutane-1-carbonitrile 65b (pale yellow solid, 227 mg, 44.6% yield). LC-MS m/z(ESI)= 322.10 [M+1]
Step 3: 4-((9-(trans-3-Cyanocyclobutyl)-7-ethyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 65)
trans-3-(7-Ethyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 65b (200 mg, 0.62 mmol) and 4-amino-3-methyl-2-fluorobenzamide intermediate 2 (538 mg, 3.20 mmol) were dissolved in N,N-dimethylformamide (3 mL), followed by notrogen purging and the addition of potassium tert-butoxide (143 mg, 1.28 mmol). The reaction mixture was stirred under nitrogen at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into 30 mL of purified water, and a large amounts of yellow solid was precipitated. The solid was collected by filtration and purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(cis-3-cyanocyclobutyl)-7-ethyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide compound 65 (white solid, 15 mg, 5.9% yield). 1H NMR (400 MHz, DMSO-d6) 68.64 (s, 1H), 8.27(s, 1H), 7.86(d 1H), 7.56-7.54(m, 1H), 7.48(s, 1H), 7.40(s, 1H), 5.14-5.09(m, 1H), 3.86-3.82(m, 2H), 3.64-3.59(m, 1H), 3.26-3.16(m, 2H), 2.75-2.68(m, 2H), 2.29(s, 3H), 1.22(t, 3H). LC-MS m/z(ESI)= 410.20 [M+1]
Example 66 2-Fluoro-4-((9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide(compound66)
0 H2 N N N O Fa N N N H
N7 Compound 66 O 0 H 0N O"N:N OH N~ N N~ 0 - Step 1 I Step 2 I Step3 CI O 1. - CI N NH CI N NH C CI N CI 7 N N
1A 66a 66b 66c
nN 0
Step 4 CI N N Step 5 H2N N NN N N N== F H
7 N 66d Compound 66
Step 1: Ethyl 2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylate (66a)
2-Isopropyl-4-methylpyridin-3-amine (21.61 g, 143.92 mmol) and ethyl 2,4 dichloropyrimidine-5-carboxylate 1A (26.51 g, 119.93 mmol) were dissolved in acetonitrile (400 mL), and potassium carbonate (33.15 g, 239.86 mmol) was added with stirring at 0 °C. The reaction mixture was gradually heated to 90 °C and stirred for 72 h. The reaction was monitored by TLC. After completion of the reaction, water and ethyl acetate were added to the reaction mixture for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the title compound ethyl 2-chloro-4-((2-isopropyl-4-methylpyridin-3 yl)amino)pyrimidine-5-carboxylate 66a (brown solid, 44.23 g, 91.95% yield). 'H NMR (400 MHz, DMSO-d6) 6 9.83 (s, 1H), 8.77 (s, 1H), 8.39 (d, 1H), 7.20 (d, 1H), 4.39 (q, 2H), 3.10-3.03 (m, 1H), 2.13 (s, 3H), 1.36 (t, 3H), 1.12 (d, 6H). LC-MS m/z(ESI)= 335.10 [M+1]
Step 2: 2-Chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid (66b)
Ethyl 2-chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylate 66a (44.23 g, 131.42 mmol) was dissolved in tetrahydrofuran/water (200 mL/200 mL), and lithium hydroxide (11.03 g, 262.84 mmol) was added thereto. The reaction mixture was stirred at room temperature for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 1-2 with 2 N hydrochloric acid, and a solid was precipitated. The reaction mixture was filtered, and the filter cake was washed with water and dried to obtain the title compound 2 chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid 66b (brown solid, 29.29 g, 72.66% yield). 'H NMR (400 MHz, DMSO-d6) 6 10.23 (s, 1H), 8.72 (s, 1H), 8.38 (d, 1H), 7.20 (d, 1H), 3.11-3.04 (m, 1H), 2.13 (s, 3H), 1.26 (d, 1H), 1.12 (d, 6H). LC-MS m/z(ESI)= 307.10 [M+1]
Step 3: 2-Chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin-8-one (66c)
2-Chloro-4-((2-isopropyl-4-methylpyridin-3-yl)amino)pyrimidine-5-carboxylic acid 66b (25.00 g, 81.5 mmol) was dissolved in tetrahydrofuran. The solution was cooled to °C, and diphenylphosphoryl azide (24.6 g, 89.5 mmol) and triethylamine (9.8 g, 97.75 mmol) were added. The reaction mixture was stirred for 1 h at room temperature, and gradually heated to 120 °C for another 2 h. The reaction was monitored by TLC. After completion of the reaction, water and ethyl acetate were added to the reaction mixture for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1/1) to obtain the title compound 2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin 8-one 66c (white solid, 18 g, 75% yield). 'H NMR (400 MHz, DMSO-d6) 6 12.06 (s, 1H), 8.59 (d, 1H), 8.33 (s, 1H), 7.35 (dd, 1H), 2.80-2.73(m, 1H), 2.05 (s, 3H), 1.09 (dd, 6H). LC-MS m/z(ESI) = 304.10 [M+1]
Step 4: 2-Chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (66d)
2-Chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7,9-dihydro-8H-purin-8-one 66c (500 mg, 1.65 mmol) was dissolved in N,N-dimethylformamide (10 mL), and cesium carbonate (536 mg, 1.65 mmol) and dimethyl sulfate (269 mg, 2.14 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted three times with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1/1) to yield the title compound 2-chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9 dihydro-8H-purin-8-one 66d (white solid, 258 mg, 49.32% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.60 (d, 1H), 8.56 (s, 1H), 7.36 (d, 1H), 3.49 (s, 3H), 2.71-2.78 (m, 1H), 2.04 (s, 3H), 1.10 (d, 3H), 1.06 (d, 3H). LC-MS m/z(ESI) = 318.10 [M+1]
Step 5: 2-Fluoro-4-((9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 66)
2-Chloro-9-(2-isopropyl-4-methylpyridin-3-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 66d (100 mg, 0.31 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (210 mg, 1.26 mmol), cesium carbonate (410 mg, 1.26 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (43 mg, 0.04 mmol) were dissolved in 1,4-dioxane (4 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 6 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated and purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 20/1), and further purified by Pre HPLC to obtain the title compound 2-fluoro-4-((9-(2-isopropyl-4-methylpyridin-3-yl) 7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 66 (white solid, 40 mg, 28.28% yield). 1H NMR (400 MHz, DMSO-d6) 68.66 (s, 1H), 8.56 (d, 1H), 8.33 (s, 1H), 7.73 (s, 0.5H), 7.69 (s, 0.5H), 7.48 (d, 1H), 7.44 (s, 1H), 7.38 (d, 1H), 7.34 (d, 1H), 3.45 (s, 3H), 2.85-2.78 (m, 1H), 2.20 (s, 3H), 2.06 (s, 3H), 1.13 (s, 3H), 1.11 (s, 3H). LC-MS m/z(ESI) = 450.20 [M+1]
Example 67 2-Fluoro-4-((9-(3-hydroxycyclopentyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 67)
0 H2 N N N
Fj[N C N N )= H
OH Compound 67
O OH OH Step 1 Step 2 Step 3 N O CI N NH HN-Boc HN-Boc NH 2 HCI
67a 67b 67c 67d OH
0
N~ OH N~N N" N>= Step 4 I Step 5 I -O Step6 I - CI N NH ~ CI N N CI N N
OH OH OH 67e 67f 67g
Step 7 N N Step 8 H 2N NN F N N F NN N
H OH H OH 67h Compound 67 Step 1: tert-Butyl (3-hydroxycyclopentyl)carbamate (67b)
tert-Butyl (3-oxocyclopentyl) carbamate 67a (10 g, 50.19 mmol) was dissolved in 60 mL of tetrahydrofuran and sodium borohydride (2.85 g, 75.28 mmol) was added slowly in an ice bath. The reaction mixture was stirred for reaction for 3 h. The reaction was monitored by TLC. After completion of the reaction, saturated NaCl solution was slowly added until no air bubbles appeared, and extracted with ethyl acetate. The organic phase was concentrated to yield the title compound tert-butyl (3 hydroxycyclopentyl)carbamate 67b (yellow oily liquid, 10.10 g, 99% yield). LC-MS m/z(ESI)= 202.20 [M+1]
Step 2: 3-Aminocyclopentan-1-ol hydrochloride (67c)
tert-Butyl (3-hydroxycyclopentyl)carbamate 67b (10.10 g, 50.18 mmol) was added to a solution of 1,4-dioxane (20 mL), and the reaction mixture was stirred at room temperature until completely dissolved, placed in an ice bath, added slowly with 4 M hydrochloric acid-1,4 dioxane solution (8.75 g, 239.87 mmol) and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated to yield the title compound 3 aminocyclopentan--ol hydrochloride 67c (black oily liquid, crude product, 6.91 g, 100% yield) which was used directly in the next step. LC-MS m/z(ESI) = 138.10 [M+1]
Step 3: Ethyl 2-chloro-4-((3-hydroxycyclopentyl)amino)pyrimidine-5-carboxylate (67d)
3-Aminocyclopentane-1-ol hydrochloride 67c (6.91 g, 50.22 mmol) and ethyl 2,4 dichloropyrimidine-5-carboxylate (13.32 g, 60.26 mmol) were added to 75 mL of acetonitrile, and after being stirred at room temperature until dissolved, the reaction mixture was added slowly with potassium carbonate (17.35 g, 125.54 mmol) and stirred for 10 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove acetonitrile, and water was added and the organics were extracted with ethyl acetate. The organic phase was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1/1) to yield the title compound ethyl 2-chloro-4 ((3-hydroxycyclopentyl)amino)pyrimidine-5-carboxylate 67d (yellow oily liquid, 7.78 g, 54% yield). LC-MS m/z(ESI) = 286.10 [M+1]
Step 4: 2-Chloro-4-((3-hydroxycyclopentyl)amino)pyrimidine-5-carboxylic acid (67e)
Ethyl 2-chloro-4-((3-hydroxycyclopentyl)amino)pyrimidine-5-carboxylate 67d (7.78 g, 27.23 mmol) was dissolved in 50 mL of tetrahydrofuran, lithium hydroxide (1.3 g, 54.46 mmol) was dissolved in 50 mL of water, an aqueous solution of lithium hydroxide was slowly added to the THF solution in an ice bath and the reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction,the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 4 with hydrochloric acid solution, and a solid was precipitated. The solid was collected by filtration, and the filtrate was extracted with ethyl acetate. The organic phase and the filter residue were combined and concentrated to obtain the title compound 2-chloro-4-((3 hydroxycyclopentyl)amino)pyrimidine-5-carboxylic acid 67e (pale yellow solid, 6.78 g, 96% yield). LC-MS m/z(ESI) = 258.10 [M+1]
Step 5: 2-Chloro-9-(3-hydroxycyclopentyl)-7,9-dihydro-8H-purin-8-one (67f)
2-Chloro-4-((3-hydroxycyclopentyl)amino)pyrimidine-5-carboxylic acid 67e (6.0 g, 23.29 mmol) was dissolved in 60 mL of N,N-dimethylacetamide, diphenylphosphoryl azide (6.41 g, 23.29 mmol) and triethylamine (2.36 g, 23.29 mmol) were slowly added in an ice bath, and the reaction mixture was stirred at room temperature for 1 h, and then stirred at 90 °C for 5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was added dropwise to 200 mL of water, and extracted with ethyl acetate. The organic phase was collected and concentrated to obtain the title compound 2-chloro-9-(3-hydroxycyclopentyl)-7,9-dihydro-8H-purin-8-one 67f (yellow oily liquid, 5.47 g, 92% yield). LC-MS m/z(ESI)= 255.10 [M+1]
Step 6: 2-Chloro-9-(3-hydroxycyclopentyl)-7-methyl-7,9-dihydro-8H-purin-8-one (67h)
2-Chloro-9-(3-hydroxycyclopentyl)-7,9-dihydro-8H-purin-8-one 67f (2.5 g, 9.82 mmol) and cesium carbonate (4.16 g, 12.76 mmol) were dissolved in 40 mL of DMF, and the reaction mixture was stirred in an ice bath for 10 min, then added slowly with dimethyl sulfate (1.24 g, 9.82 mmol) and reacted for 1 h. The reaction was monitored by TLC. After completion of the reaction, 100 mL of water was added and the organics were extracted with ethyl acetate, and the organic phase was collected, concentrated and purified by column chromatography to yield the title compound 2-chloro-9-(3 hydroxycyclopentyl)-7-methyl-7,9-dihydro-8H-purin-8-one 67g (orange solid, 900 mg, 34% yield). H NMR (400 MHz, DMSO-d6) 6 8.35 (s, 1H), 4.90 (d, 1H), 4.71-4.62 (m, 1H), 4.16 4.08 (m, 1H), 3.35 (s, 3H), 2.29-2.23 (m, 1H), 2.17-2.10(m, 2H), 1. 92-1.77 (m, 3H). LC-MS m/z(ESI)= 269.10 [M+1]
Step 7: 2-Fluoro-4-((9-(3-hydroxycyclopentyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzonitrile (50h)
2-Chloro-9-(3-hydroxycyclopentyl)-7-methyl-7,9-dihydro-8H-purin-8-one 67g (600 mg, 2.23 mmol), 4-amino-2-fluoro-5-methylbenzonitrile 2A (335 mg, 2.23 mmol), cesium carbonate (2.18 g, 6.70 mmol), and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (303 mg, 0.34 mmol) were dissolved in 1,4-dioxane (10 mL), and the reaction mixture was purged with nitrogen and stirred at 110 °C for 6 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 20/1) to obtain the title compound 2-fluoro-4-((9-(3-hydroxycyclopentyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzonitrile 67h (pale yellow solid, 460 mg, 54% yield). H NMR (400 MHz, DMSO-d6) 6 8.74 (d, 1H), 8.25 (d, 1H), 8.20-8.15 (m, 1H), 7.66 (d, 1H), 5.05-4.96 (m, 0.5H), 4.83 (d, 0.5H), 4.73-4.64 (m, 0.5H), 4.66 (s, 0.5H), 4.13 4.08 (m, 1H), 3.34 (d, 3H), 2.31 (s, 3H), 2.15-2.09 (m, 3H), 1.96-1.75 (m, 3H). LC-MS m/z(ESI)= 383.20 [M+1]
Step 8: 2-Fluoro-4-((9-(3-hydroxycyclopentyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 67)
2-Fluoro-4-((9-(3-hydroxycyclopentyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzonitrile 67h (400 mg, 1.05 mmol) and potassium carbonate (43.4 mg, 313.81 mmol) were dissolved in 20 mL of dimethyl sulfoxide, and hydrogen peroxide (1.05 mL, 10.46 mmol) was slowly added with stirring at room temperature. The reaction mixture was stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by preparative medium pressure liquid chromatography, followed by Pre HPLC to obtain the title compound 2-fluoro-4-((9-(3-hydroxycyclopentyl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 67, which was then resolved by SFC to yield compound 67-1 (white solid, 70 mg, 17% yield, dr = 99%) and compound 67-2 (white solid, 58 mg, 14% yield, dr = 99%). (OZ), mobile phase: C02/isopropanol= 70/30; column temperature: 35 °C; column pressure: 80 bar; flow rate: 1 mL/min; detector signal channel: 220 nm@4 nm; diode array detector at a wavelength of 200-400 nm.
Compound 67-1: H NMR (400 MHz, DMSO-d6) 6 8.48 (s, 1H), 8.19 (s, 1H), 7.88 (d, 1H), 7.54 (d, 1H), 7.47 (s, 1H), 7.37 (d, 1H), 4.86 (d, 1H), 4.72-4.63 (m, 1H), 4.14-4.06 (m, 1H), 3.32(s, 3H), 2.42-2.32 (m, 1H), 2.28 (s, 3H), 2.16-2.10 (m, 2H), 1.93-1.76 (m,3H). LC-MS m/z(ESI)= 400.20 [M+1] Compound 67-2: 1H NMR (400 MHz, DMSO-d6) 68.51 (s, 1H), 8.18 (s, 1H), 7.86 (d, 1H), ,7.54 (d, 1H), 7.46 (s, 1H), 7.38 (d,1H), 5.03-4.94 (m, 1H), 4.64(d, 1H), 4.35 (s,H), 3.32(s,3H), 2.38-2.31(m,1H), 2.28 (s, 3H), 2.15-2.03(m,3H)1.85-1.80(m, 1H), 1.62-1.57(m, 1H). LC-MS m/z(ESI)= 400.20 [M+1]
Example 68 4-((9-Cyclohexyl-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5 methylbenzamide (compound 68)
0
H2 N N NO F 1: N N N H
Compound 68 0 0 N OH Step N O' Step2 N Step3 N CI NN CI N NH CI N NH CI N CI
IA 68a 68b
H I0 NN NHN~ ~ N I -O Step 4 IC CIl - N>= N>= -O Step 4 H2N FC bCl N N H-,''N>= N I H
68c 68d Compound 68
Step 1: Ethyl 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylate (68a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5.0 g, 22.6 mmol) and potassium carbonate (6.2 g, 45.2 mmol) were dissolved in acetonitrile (50 mL), and cyclohexylamine (2.2 g, 22.6 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate., and was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)= 10:1) to obtain the title compound ethyl 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylate 68a (white solid, 4.1 g, 64% yield). 1H NMR (400 MHz, Chloroform-d) 6 8.64 (s, 1H), 8.53-8.23 (m, 1H), 4.35 (q, 2H), 4.25-4.05 (m, 1H), 2.01-1.95 (m, 2H), 1.77-1.72(m, 2H), 1.64-1.61 (m,1H), 1.50-1.41 (m, 3H), 1.39 (t, 3H), 1.35-1.18 (m, 2H). LC-MS m/z(ESI) = 284.10 [M+1]
Step 2: 2-Chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid (68b)
Ethyl 2-chloro-4-(cyclohexylamino)pyrimidine-5-carboxylate 68a (4.1 g, 14.4 mmol) was dissolved in tetrahydrofuran/water (20 mL/20 mL), and lithium hydroxide (691 mg, 28.8 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 2-chloro-4 (cyclohexylamino)pyrimidine-5-carboxylic acid 68b (white solid, 3.0 g, 81% yield). H NMR (400 MHz, DMSO-d6) 6 13.77(s, 1H), 8.76-8.45 (m, 2H), 4.01-3.92 (m, 1H), 2.05-1.79 (m, 2H), 1.79-1.65 (m, 2H), 1.60-1.54 (m, 1H), 1.42-1.19 (m, 5H). LC-MS m/z(ESI) = 256.10 [M+1]
Step 3: 2-Chloro-9-cyclohexyl-7,9-dihydro-8H-purin-8-one (68c)
2-Chloro-4-(cyclohexylamino)pyrimidine-5-carboxylic acid 68b (3.0 g, 11.7 mmol) was dissolved in dimethylacetamide (30 mL), and triethylamine (1.18 g, 11.7 mmol) and diphenylphosphoryl azide (3.22 g, 11.7 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:10) to obtain the title compound 2-chloro-9-cyclohexyl-7,9-dihydro 8H-purin-8-one 68c (white solid, 2.2 g, 74% yield). H NMR (400 MHz, DMSO-d6) 6 11.60 (s, 1H), 8.12 (s, 1H), 4.19-4.11 (m, 1H), 1.92 1.57 (m, 6H), 1.52-0.93 (m, 4H). LC-MS m/z(ESI) = 253.10 [M+1]
Step 4: 2-Chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one (68d)
2-Chloro-9-cyclohexyl-7,9-dihydro-8H-purin-8-one 68c (2.2 g, 8.7 mmol) was dissolved in dimethylformamide (15 mL), and dimethyl sulfate (1.1 g, 8.7 mmol) and cesium carbonate (4.25 g, 13 mmol) were added at 0 °C. The reaction mixture was stirred for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to give the title compound 2-chloro-9-cyclohexyl-7-methyl-7,9 dihydro-8H-purin-8-one 68d (yellow solid, 1.1 g, 48% yield). H NMR (400 MHz, DMSO-d6) 6 8.34 (s, 1H), 4.22-4.41 (m, 1H), 3.35 (s, 3H), 2.22 2.12 (m, 2H), 1.91-1.60 (m, 5H), 1.43-1.33 (m, 2H), 1.27-1.14 (m, 1H). LC-MS m/z(ESI)= 267.10 [M+1]
Step 5: 4-((9-Cyclohexyl-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5 methylbenzamide (compound 68)
2-Chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one 68d (200 mg, 0.79 mmol), 4-amino-3-methyl-2-fluorobenzamide (292 mg, 1.74 mmol), cesium carbonate (515 mg, 1.58 mmol) and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy 2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (72 mg, 0.079 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 35/1) to give the title compound 4-((9-cyclohexyl 7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 68 (white solid, 80 mg, 25 % yield). 'H NMR (400 MHz, DMSO-d6) 6 8.48 (s, 1H), 8.18 (s, 1H), 8.01 (d, 1H), 7.54 (d, 1H), 7.42 (d, 2H), 4.23-4.14 (m, 1H), 3.32 (s, 3H), 2.30 (s, 3H), 2.29-2.15 (m, 2H), 1.85 1.67 (m, 5H), 1.55-1.07 (m, 3H). LC-MS m/z(ESI)= 399.20 [M+1]
Example 69 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 69)
0 H 2N N NO F N N N H 0 Compound 69 0 0
Step N 0 ' O Step 2 Step 3 N N 0' - CI N NH CI N NH CI N CI
1A 69a 69b
H 0 I O step 4 C Step 5 H2N N IN IN -Q-p4':N>=0 Step5 :>== CI NIN CI NN F 1:N INN H
69c 69d Compound 69
Step 1: Ethyl 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylate (69a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5.0 g, 22.6 mmol) and potassium carbonate (6.2 g, 45.2 mmol) were dissolved in acetonitrile (50 mL), and tetrahydro 2H-pyran-3-amine (2.3 g, 22.6 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. water was added and the organics were extracted three times with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v) = 10:1) followed by concentration to obtain the title compound ethyl 2-chloro-4 ((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylate 69a (white solid, 4.1 g, 64% yield).
H NMR (400 MHz, Chloroform-d) 6 8.69 (s, 1H), 8.68 (s,1H), 4.37(q, 2H),4.34-4.24 (m, 1H), 3.89-3.84 (m, 1H), 3.77-3.66 (m, 2H), 3.58-3.54 (m, 1H), 2.01-1.95 (m, 1H), 1.86-1.65 (m, 2H), 1.69-1.63 (m, 1H), 1.40 (t, 3H). LC-MS m/z(ESI) = 286.10 [M+1]
Step 2: 2-Chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid (69b)
Ethyl 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylate 69a (4.1 g, 14.3 mmol) was dissolved in tetrahydrofuran/water (20 mL/20 mL), and lithium hydroxide (686 mg, 28.6 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to give the title compound 2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid 69b (white solid, 3.5 g, 95% yield). 'H NMR (400 MHz, DMSO-d6) 6 13.83 (s, 1H), 8.75 (d, 1H), 8.59 (s, 1H), 4.14-4.07 (m, 1H), 3.76-3.72 (m, 1H), 3.59 (t, 1H), 3.47-3.42 (m, 2H), 1.93-1.86 (m, 1H), 1.79 1.62 (m, 2H), 1.58-1.49(m, 1H). LC-MS m/z(ESI) = 258.10 [M+1]
Step 3: 2-Chloro-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one (69c)
2-Chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylic acid 69b (3.5 g, 13.6 mmol) was dissolved in dimethylacetamide (30 mL), and triethylamine (1.37 g, 13.6 mmol) and diphenylphosphoryl azide (3.74 g, 13.6 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:10) to obtain the title compound 2-chloro-9 (tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one 69c (white solid, 2.2 g, 63% yield). H NMR (400 MHz, DMSO-d6) 6 11.67 (s, 1H), 8.13 (s, 1H), 4.29-4.21 (m, 1H), 3.86 3.77 (m, 2H), 3.37-3.29 (m, 2H), 2.04-1.39 (m, 4H). LC-MS m/z(ESI)= 255.10 [M+1]
Step 4: 2-Chloro-7-methyl-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one (69d)
2-Chloro-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one 69c (2.2 g, 8.7 mmol) was dissolved in dimethylformamide (15 mL), and dimethyl sulfate (1.1 g, 8.7 mmol) and cesium carbonate (4.25 g, 13 mmol) were added at 0 °C. The reaction mixture was stirred for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to give the title compound 2-chloro-9-cyclohexyl-7 methyl-7,9-dihydro-8H-purin-8-one 69d (yellow solid, 1.7 g, 73% yield). H NMR (400 MHz, DMSO-d6) 6 8.36 (s, 1H), 4.35-4.23 (m, 1H), 3.91-3.77 (m, 2H),
3.35 (s, 3H), 3.34-3.33(m, 2H), 2.50-2.44 (m, 1H), 1.98-1.89 (m, 1H), 1.83-1.62 (m, 2H). LC-MS m/z(ESI) = 269.10 [M+1]
Step 5: 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 69)
2-Chloro-7-methyl-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one 69d (200 mg, 0.75 mmol), 4-amino-3-methyl-2-fluorobenzamide intermediate 2 (402 mg, 1.50 mmol), cesium carbonate (489 mg, 1.50 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (68 mg, 0.075 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 2-fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydro-2H-pyran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamide compound 69, which was then resolved by SFC to yield compound 69-1 (white solid, 32 mg, 11% yield, RT = 12.47 min) and compound 69 2 (white solid, 33 mg, 11% yield, RT = 14.31 min). (OZ), mobile phase: C02/ethanol = 65/35; column temperature: 35 °C; column pressure: 80 bar; flow rate: 1 mL/min; detector signal channel: 215 nm@4.8 nm; diode array detector at a wavelength of 200 400 nm. 1H NMR (400 MHz, DMSO-d6) 6 8.54 (s, 1H), 8.20 (s, 1H), 7.97 (d, 1H), 7.56 (d, 1H), 7.43 (d, 2H), 4.36-4.28 (m, 1H), 3.99 (t, 1H), 3.94-3.74 (m, 2H), 3.37(d, 1H), 3.32 (s, 3H), 2.62-2.52 (m, 1H), 2.30 (s, 3H), 1.93 (d, 1H), 1.86-1.59 (m, 2H). LC-MS m/z (ESI) = 401.20 [M+1]
Example 70 2-Fluoro-4-((9-(cis-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 70)
0 H 2N N NO F N'1N N H
0 Compound 70
0 0 Step1 N N O0 -O Step2 N OH Step3
CI N CI CI N NH y IA 70a 70b
H 0
1. >zO Step 4 1 >= tp~ NN N CI N Ste CI N Step 5 HFN N N H
0- 0-0 70c 70d Compound 70
Step 1: Ethyl 2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate (70a)
Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1A (5.13 g, 23.22 mmol) was dissolved in acetonitrile (20 mL), and potassium carbonate (6.42 g, 46.44 mmol) was added with stirring at 0 °C, followed by the slow dropwise addition of a solution of cis-4 methoxycyclohexan-1-amine (2.00 g, 15.48 mmol) in acetonitrile (10 mL). The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 2:1) to obtain the title compound ethyl 2-chloro 4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate 70a (white solid, 2.00 g, 41.26% yield). LC-MS m/z(ESI) = 314.10 [M+1]
Step 2: 2-Chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid (70b)
Ethyl 2-chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate 70a (2.00 g, 6.37 mmol) was dissolved in tetrahydrofuran/water (15 mL/15 mL), and lithium hydroxide monohydrate (0.80 g, 19.12 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to 3-4 with 2 N HCl, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to yield the title compound 2-chloro-4 ((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid 70b (white solid, 1.67 g, 91.70% yield). LC-MS m/z(ESI) = 286.10 [M+1]
Step 3: 2-Chloro-9-(cis-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one (70c)
2-Chloro-4-((cis-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid 70b (1.57 g, 5.49 mmol) was dissolved in N,N-dimethylacetamide (16 mL), and triethylamine (0.76 mL, 5.49 mmol) and diphenylphosphoryl azide (1.18 mL, 5.49 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with dichloromethane. The organic layer was concentrated to give the title compound 2-chloro-9-(cis-4 methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one 70c (white solid, 1.70 g, crude product, 109.47% yield). LC-MS m/z(ESI)= 283.00 [M+1]
Step 4: 2-Chloro-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (70d)
2-Chloro-9-(cis-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one 70c (1.70 g, 6.01 mmol) was dissolved in N,N-dimethylformamide (20 mL), and dimethyl sulfate (0.57 mL, 6.01 mmol) and cesium carbonate (3.92 g, 12.03 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred, and a white solid was precipitated. The solid was collected by filtration and washed 3 times with water and petroleum ether to give the title compound 2-chloro-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H purin-8-one 70d (white solid, 1.2 g, 67.25% yield). LC-MS m/z(ESI)= 297.10 [M+1]
Step 5: 2-Fluoro-4-((9-(cis-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 70)
2-Chloro-9-(cis-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 70d (0.25 g, 0.84 mmol) was dissolved in 1,4-dioxane (8 mL), and 4-amino-2-fluoro-5 methylbenzamide intermediate 2 (0.57 g, 3.37 mmol), cesium carbonate (0.41 g, 1.26 mmol) and BrettPhos-G3-Pd (0.08 g, 0.08 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 30/1) to obtain the title compound 2-fluoro-4-((9-(cis-4-methoxycyclohexyl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 70 (white solid, 95 mg, 26.39% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.47 (s, 1H), 8.16 (s, 1H), 7.82 (d, 1H), 7.55 (d, 1H), 7.44 (s, 1H), 7.35 (d, 1H), 4.26-4.18(m, 1H), 3.42-3.40 (m, 1H), 3.31 (s, 3H), 3.21 (s, 3H), 2.49-2.39 (m, 2H), 2.28 (s, 3H), 2.00 (d, 2H), 1.65-1.37 (m, 4H). LC-MS m/z(ESI) = 429.20 [M+1]
Example 71 2-Fluoro-4-((9-(trans-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin 2-yl)amino)-5-methylbenzamide (compound 71)
H 2N N
F N<N N H
0 Compound 71 0 0p N OH 0 Step 1 N H Step 2 Step 3 N O"' N"CNNH C -CI N NHj CI N CI
ON O IA 71a 71b
H 0
CIN N. >= Step 4 CI N. >o N>N - Step 5HN F N N" N >= N
0-b- 0- 0 71c 71d Compound 71
Step 1: Ethyl 2-chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate (71a)
Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1A (5.13 g, 23.22 mmol) was dissolved in acetonitrile (20 mL), and potassium carbonate (6.42 g, 46.44 mmol) was added with stirring at 0 °C, followed by the slow dropwise addition of a solution of trans-4 methoxycyclohexan-1-amine (2.00 g, 15.48 mmol) in acetonitrile (10 mL). The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 2:1) to get the title compound ethyl 2-chloro-4 ((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate 71a (white solid, 2.00 g, 41.26% yield). LC-MS m/z(ESI) = 314.10 [M+1]
Step 2: 2-Chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid (71b)
Ethyl 2-chloro-4-(((1R,4R)-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylate 71a (2.00 g, 6.37 mmol) was dissolved in tetrahydrofuran/water (15 mL/15 mL), and lithium hydroxide monohydrate (0.80 g, 19.12 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH adjusted to about 3-4 with 2 N HCl, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to get the title compound 2-chloro-4 ((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid 71b (white solid,
1.67 g, 91.70% yield). LC-MS m/z(ESI) = 286.10 [M+1]
Step 3: 2-Chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one (71c)
2-Chloro-4-((trans-4-methoxycyclohexyl)amino)pyrimidine-5-carboxylic acid 71b (1.57 g, 5.49 mmol) was dissolved in N,N-dimethylacetamide (16 mL), and triethylamine (0.76 mL, 5.49 mmol) and diphenylphosphoryl azide (1.18 mL, 5.49 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, Water was added and the organics were extracted with dichloromethane. The organic layer was concentrated to give the title compound 2-chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one 71c (white solid, 1.70 g, crude product, 109.47% yield). 'H NMR (400 MHz, DMSO-d6) 6 11.61 (s, 1H), 8.11 (s, 1H), 4.20-4.10 (m, 1H), 3.25 (s, 3H), 3.22-3.14 (m, 1H), 2.30-2.18 (m, 2H), 2.15-2.06 (m, 2H), 1.81-1.73 (d, 2H), 1.31-1.18 (m, 2H). LC-MS m/z(ESI)= 283.00 [M+1]
Step 4: 2-Chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (71d)
2-Chloro-9-(trans-4-methoxycyclohexyl)-7,9-dihydro-8H-purin-8-one 71c (1.70 g, 6.01 mmol) was dissolved in N,N-dimethylformamide (20 mL), and dimethyl sulfate (0.57 mL, 6.01 mmol) and cesium carbonate (3.92 g, 12.03 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred, and a white solid was precipitated. The solid was collected by filtration and washed 3 times with water and petroleum ether to obtain the title compound 2-chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-7,9 dihydro-8H-purin-8-one 71d (white solid, 1.2 g, 67.25% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.34 (s, 1H), 4.26-4.15 (m, 1H), 3.35 (s, 3H), 3.27 (s, 3H), 3.25-3.16 (m, 1H), 2.31-2.19 (m, 2H), 2.15-2.08 (m, 2H), 1.83-1.75 (m, 2H), 1.34-1.21 (m, 2H). LC-MS m/z(ESI)= 297.10 [M+1]
Step 5: 2-Fluoro-4-((9-(trans-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin 2-yl)amino)-5-methylbenzamide (compound 71)
2-Chloro-9-(trans-4-methoxycyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 71d (0.25 g, 0.84 mmol) was dissolved in 1,4-dioxane (8 mL), and 4-amino-2-fluoro-5 methylbenzamide (0.57 g, 3.37 mmol), cesium carbonate (0.41 g, 1.26 mmol) and BrettPhos-G3-Pd (0.08 g, 0.08 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 30/1) to obtain the title compound 2-fluoro-4-((9-(trans-4-methoxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro
7H-purin-2-yl)amino)-5-methylbenzamide compound 71 (white solid, 130 mg, 36.01% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.48 (s, 1H), 8.18 (s, 1H), 7.98 (d, 1H), 7.56 (d, 1H), 7.46 (s, 1H), 7.39 (s, 1H), 4.26-4.16 (m, 1H), 3.34 (s, 3H), 3.28 (s, 3H), 3.25-3.16 (m, 1H), 2.42-2.33 (m, 2H), 2.30 (s, 3H), 2.13 (d, 2H), 1.79 (d, 2H), 1.34-1.19 (m, 2H). LC-MS m/z(ESI)= 429.20 [M+1]
Example 72 2-Fluoro-4-((9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide(compound72)
0I
H 2N N N NO F N N N H
OH Compound 72 0 0 O OH Step N '.O-"' Step2 N H Step3 N ' | CI N H CI N NH CI N CI
OH OH OH 1A 72a 72b
H 0/ N Stp4 N N >=oH 2N N N I >=O Step 4 1 -O Step 5 H2NN O CI N NC' H
OH OH OH 72c 72d Compound 72
Step 1: Ethyl 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5 carboxylate (72a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5.0 g, 22.6 mmol) and potassium carbonate (6.2 g, 45.2 mmol) were dissolved in acetonitrile (50 mL), and cis-4-amino 1-methylcyclohexan-1-ol (2.9 g, 22.6 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. water was added and the organics were extracted three times with ethyl acetate., and the organic phase was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n hexane/ethyl acetate (v/v) = 10:1) followed by concentration to obtain the title compound ethyl 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5 carboxylate 72a (white solid, 4.1 g, 58% yield). 1H NMR (400 MHz, Chloroform-d) 6 8.65 (s, 1H), 8.37 (d, 1H), 4.35 (q, 2H), 4.09
4.04 (m, 1H), 1.94-1.82 (m, 2H), 1.82-1.45 (m, 6H), 1.38 (t, 3H), 1.28 (s, 3H). LC-MS m/z(ESI) = 314.10 [M+1]
Step 2: 2-Chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (72b)
Ethyl 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5 carboxylate 72a (4.1 g, 13.1 mmol) was dissolved in tetrahydrofuran/water (20 mL/20 mL), and lithium hydroxide (629 mg, 26.2 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to give the title compound 2-chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 72b (white solid, 3.5 g, 93% yield). 'H NMR (400 MHz, DMSO-d6) 6 13.77(br, 1H), 8.56 (s, 1H), 8.49 (d, 1H), 3.93-3.83 (m, 2H), 1.78-1.50 (m, 6H), 1.43-1.40(m, 2H), 1.11 (s, 3H). LC-MS m/z(ESI) = 286.10 [M+1]
Step 3: 2-Chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (72c)
2-Chloro-4-((cis-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 72b (3.5 g, 12.2 mmol) was dissolved in dimethylacetamide (30 mL), and triethylamine (1.24 g, 12.2 mmol) and diphenylphosphoryl azide (3.36 g, 12.2 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:10) to obtain the title compound 2-chloro-9 (cis-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 72c (white solid, 1.4 g, 41% yield). H NMR (400 MHz, DMSO-d) 6 11.58 (s, 1H), 8.11 (d, 1H), 4.22(s, 1H), 4.22-4.02 (m, 1H), 2.65-2.56 (m, 2H), 1.83-1.60 (m, 2H), 1.47-1.43 (m, 4H), 1.18 (s, 3H). LC-MS m/z(ESI)= 283.10 [M+1]
Step 4: 2-Chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8 one (72d)
2-Chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 72c (1.4 g, 4.9 mmol) was dissolved in dimethylformamide (15 mL), and dimethyl sulfate (618 mg, 4.9 mmol) and cesium carbonate (3.2 g, 9.8 mmol) were added at 0 °C. The reaction mixture was stirred for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to give the title compound 2-chloro-9-(cis-4-hydroxy 4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 72d (yellow solid, 502 mg, 34% yield). LC-MS m/z(ESI)= 297.10 [M+1]
Step 5: 2-Fluoro-4-((9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 72)
2-Chloro-9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8-purin-8-one 72d (250 mg, 0.84 mmol), 4-amino-3-methyl-2-fluorobenzamide intermediate 2 (282 mg, 1.68 mmol), cesium carbonate (446 mg, 1.68 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (76 mg, 0.084 mmol) were dissolved in dioxane (4 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 2-fluoro-4-((9-(cis-4-hydroxy-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide compound 72 (white solid, 80 mg, 22 % yield). 1H NMR (400 MHz, DMSO-d6) 6 8.40 (s, 1H), 8.15 (s, 1H), 7.92 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.35 (d, 1H), 4.20-4.06 (m, 1H), 4.02 (s, 1H), 3.31 (s, 3H), 2.78-2.57 (m, 2H), 2.28 (s, 3H), 1.68 (d, 2H), 1.49-1.40 (m, 4H), 1.15 (s, 3H). LC-MS m/z (ESI) = 429.20 [M+1]
Example 73 2-Fluoro-4-((9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-5-methylbenzamide(compound73)
0
H2 N N NO F N<N N H
oH Compound 73 0 0 Step N O O Step2 N Step3 N O' I - CI NH CN N NH CI N CI
1A 73a 73b
H 0
CI -O Step 4 N CIO Step 5 H2N N O CI N CINN== F NN N H
OH OH OH 73d Compound 73 73c Step 1: Ethyl 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5 carboxylate (73a)
Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1A (5.00 g, 30.18 mmol) was dissolved in acetonitrile (30 mL), and potassium carbonate (12.51 g, 90.55 mmol) was added with stirring at 0 °C, followed by the slow dropwise addition of a solution of trans-4-amino 1-methylcyclohexanol hydrochloride (10.01 g, 45.27 mmol) in acetonitrile (10 mL). The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 2:1) to obtain the title compound ethyl 2-chloro 4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate 73a (white solid, 4.70 g, 49.63% yield). H NMR (400 MHz, Chloroform-d) 6 8.66 (s, 1H), 4.36 (q, 2H), 4.30-4.21 (m, 1H), 2.10-2.00 (m, 2H), 1.74-1.63 (m, 5H), 1.62-1.50 (m, 3H), 1.40 (t, 3H), 1.31 (s, 3H). LC-MS m/z(ESI) = 314.10 [M+1]
Step 2: 2-Chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (73b)
Ethyl 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5 carboxylate 73a (4.70 g, 14.98 mmol) was dissolved in tetrahydrofuran/water (40 mL/20 mL), and lithium hydroxide monohydrate (1.89 g, 44.94 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to about 3-4 with 2 N HCl, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to give the title compound 2-chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5 carboxylic acid 73b (white solid, 4.20 g, 98.15% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.68 (d, 1H), 8.56 (s, 1H), 4.02 (s, 1H), 3.16 (s, 1H), 1.91-1.83 (m, 2H), 1.51-1.45 (m, 6H), 1.15 (s, 3H). LC-MS m/z(ESI) = 314.10 [M+1]
Step 3: 2-Chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (73c)
2-Chloro-4-((trans-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 73b (4.34 g, 15.19 mmol) was dissolved in N,N-dimethylacetamide (30 mL), and triethylamine (2.11 mL, 15.19 mmol) and diphenylphosphoryl azide (3.27 mL, 5.19 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a white solid was precipitated. The reaction mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to obtain the title compound 2-chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H purin-8-one 73c (white solid, 1.82 g, 42.36% yield). LC-MS m/z(ESI)= 283.00 [M+1]
Step 4: 2-Chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8 one (73d)
2-Chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 73c (1.80 g, 6.37 mmol) was dissolved in N,N-dimethylformamide (20 mL), and dimethyl sulfate (0.60 mL, 6.37 mmol) and cesium carbonate (3.11 g, 9.55 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred, and a white solid was precipitated. The solid was collected by filtration and washed 3 times with water and petroleum ether to give the title compound 2-chloro-9-(trans-4-hydroxy-4 methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 73d (white solid, 0.80 g, 42.34% yield). H NMR (400 MHz, DMSO-d6) 6 8.35 (s, 1H), 4.48 (s, 1H), 4.23-4.12 (m, 1H), 3.35 (s, 3H), 2.42-2.25 (m, 2H), 1.65 (d, 4H), 1.58-1.47 (m, 2H), 1.26 (s, 3H). LC-MS m/z(ESI)= 297.10 [M+1]
Step 5: 2-Fluoro-4-((9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-5-methylbenzamide (compound 73)
2-Chloro-9-(trans-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8 one 73d (0.30 g, 1.01 mmol) was dissolved in 1,4-dioxane (8 mL), and 4-amino-2 fluoro-5-methylbenzamide intermediate 2 (0.37 g, 2.22 mmol), cesium carbonate (0.49 g, 1.52 mmol) and BrettPhos-G3-Pd (0.09 g, 0.10 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 30/1) to obtain the title compound 2-fluoro-4-((9-(trans-4-hydroxy-4-methylcyclohexyl)-7 methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 73 (pale yellow solid, 23 mg, 5.31% yield). H NMR (400 MHz, DMSO-d6) 6 8.56 (s, 1H), 8.17 (s, 1H), 7.72 (d, 1H), 7.55 (d, 1H), 7.45 (s, 1H), 7.34 (d, 1H), 4.41 (s, 1H), 4.16 (tt, 1H), 2.36-2.29 (m, 2H), 2.27 (s, 3H), 1.63 (d, 4H), 1.49 (td, 2H), 1.14 (s, 3H). LC-MS m/z(ESI)= 429.20 [M+1]
Example 74 4-((9-(cis-3-Cyanocyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 74)
0 H 2N - JNNj N
F N N N H
CN Compound 74
0 0 0 H
SN Step I Step2 Step 3 OH Step 4 NA- NO S HS N NH S N NHS N N
COOH CONH2 CN CN CN
74a 74b 74c 74d 113e
I / 0
step 5 O Step 0 >==O Step 7 H 2N StepS - N tep6 - NSINN NA- o NH C N CN CN
74f 74g Compound 74
Step 1: Ethyl 4-((cis-3-carbamoylcyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate (74b)
The starting material cis-3-((5-(ethoxycarbonyl)-2-(methylthio)pyrimidine-4 yl)amino)cyclobutane--carboxylic acid 74a (13.0 g, 41.8 mmol) was added to dichloromethane (130 mL), and N,N,NN-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (19.1 g, 50.2 mmol) and triethylamine (16.9 g, 167.2 mmol) were added. The mixture was stirred at room temperature for 10 min, and then ammonium chloride (6.7 g, 125.4 mmol) was added. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, 50 mL of saturated aqueous sodium bicarbonate were added to the reaction mixture and the organics were with dichloromethane, and the organic layer was concentrated, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude title compound ethyl 4-((cis-3-carbamoylcyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 74b (yellow viscous liquid, 17.5 g, 134.6% yield). LC-MS m/z(ESI)= 311.20 [M+1]
Step 2: Ethyl 4-((cis-3-cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate (74c)
Ethyl 4-((cis-3-carbamoylcyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 74b (17.5 g, 5.64 mmol) was added to dichloromethane (130 mL), and pyridine (13.2 g, 167.2 mmol) and trifluoroacetic anhydride (26.3 g, 125.4 mmol) were added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the pH of reaction mixture was adjusted to 3 with 2 N hydrochloric acid and extracted with dichloromethane, and was dried over anhydrous sodium sulfate and concentrated under reduced pressure to get a crude product, which was then purified by column chromatography (petroleum ether/ethyl acetate (v/v) = 5:1) to obtain the title compound ethyl 4-((cis-3 cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate74c(whitesolid,3.5 g, 21.2 % yield). H NMR (400 MHz, DMSO-d6) 6 8.65 (s, 1H), 8.46 (d, 1H), 4.70-4.61 (in, 1H), 4.37 4.31 (in, 2H), 2.94-2.84 (in, 3H), 2.52 (s, 3H), 2.48-2.38 (in, 2H), 1.38(t, 3H). LC-MS m/z(ESI)= 293.20 [M+1]
Step 3: 4-((cis-3-Cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid (74d)
Ethyl 4-((cis-3-cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylate 74c (3.5 g, 11.9 mmol) was dissolved in a mixture of tetrahydrofuran (20 mL) and water (20 mL), and lithium hydroxide (691 mg, 28.8 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to affordthetitle compound 4-((cis-3-cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 74d (white solid, 3.0 g, 94.6% yield). 'H NMR (400 MHz, DMSO-d6) 6 13.30 (br, 1H) 8.58 (d, 1H), 8.53 (s, 1H), 4.66-4.56 (m, 1H), 3.18-3.09 (m, 1H), 2.74-2.65(m, 2 H), 2.47 (s, 3H), 2.48-2.40 (m, 2H). LC-MS m/z(ESI)= 265.10 [M+1]
Step 4: cis-3-(2-(Methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile (74e)
4-((cis-3-Cyanocyclobutyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 74d (3.0 g, 11.3 mmol) was dissolved in N,N-dimethylformamide (30 mL), and triethylamine (1.18 g, 11.7 mmol) and diphenylphosphoryl azide (3.22 g, 11.7 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 5:1 to 1:10) to obtain the title compound cis-3-(2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 74e (white solid, 2.7 g, 90.9% yield). 'H NMR (400 MHz, DMSO-d6) 6 11.35 (s, 1H), 8.11 (s, 1H), 4.90-4.02 (m, 1H), 3.37 3.26 (m, 2H), 3.25-3.18 (m, 1H), 2.78-2.66 (m, 2H), 2.58 (s, 3H). LC-MS m/z(ESI)= 262.10 [M+1]
Step 5: cis-3-(7-Methyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (74f)
cis-3-(2-(Methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 74e (500 mg, 19.1 mmol) was dissolved in dimethylformamide (5 mL), and dimethyl sulfate (2.4 g, 19.1 mmol) and cesium carbonate (12.5 g, 38.2 mmol) were added at °C. The reaction mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, then 50 mL of water was added, and a solid was precipitated. The solid was collected by filtration and dried by rotary evaporation under reduced pressure to get the title compound cis-3-(7-methyl-2-(methylthio)-8-oxo-7,8 dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 74f (white solid, 457 mg, 86.7% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.61 (s, 1H), 4.90-4.02 (m, 1H), 3.34(s, 3H), 3.30 3.23(m, 2H), 3.25-3.18(m, 1H), 2.89(s, 3H), 2.76-2.71(m, 2H). LC-MS m/z (ESI)= 276.10 [M+1]
Step 6: cis-3-(7-Methyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (74g)
cis-3-(7-Methyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 74f (457 mg, 1.66 mmol) was dissolved in methanol (10 mL), and 2 mL of purified water was added, followed by the addition of potassium peroxymonosulfate (1.02 g, 1.66 mmol). The reaction mixture was heated to 60 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove methanol, and 5 mL of purified water was added. The resulting mixture was extracted 3 times with dichloromethane (20 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent and thus to provide the title compound cis-3-(7-methyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro 9H-purin-9-yl)cyclobutane--carbonitrile 74g (pale yellow solid, 197 mg, 38.6% yield). LC-MS m/z(ESI)= 308.10 [M+1]
Step 7: 4-((9-(cis-3-Cyanocyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 74)
cis-3-(7-Methyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 74g (197 mg, 0.64 mmol) and 4-amino-3-methyl-2-fluorobenzamide (538 mg, 3.20 mmol) were dissolved in N,N-dimethylformamide (3 mL), followed by nitrogen purging and the addition of potassium tert-butoxide (143 mg, 1.28 mmol). The reaction mixture was stirred under nitrogen at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into 30 mL of purified water, and a large amounts of yellow solid was precipitated. The solid was collected by filtration and purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(cis-3-cyanocyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide compound 74 (white solid, 25 mg, 10.0 % yield). 1H NMR (400 MHz, DMSO-d6) 68.63 (s, 1H), 8.20(s, 1H), 7.87(d 1H), 7.56-7.54(m, 1H), 7.48(s, 1H), 7.40(s, 1H), 5.14-5.09(m, 1H), 3.64-3.59(m, 1H), 3.29(s, 3H), 3.26 3.16(m, 2H), 2.75-2.68(m, 2H), 2.30(s, 3H). LC-MS m/z(ESI)= 396.20 [M+1]
Example 75 (S)-2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamiderile (compound 75)
H 2N N~ I N )= F N N - H S
Compound 75
0 0 0 Step 1 N 0- Step 2 N OH Step 3
CI [ N NH CI N NH CI' N CI S
1A 75a 75b
H 0 N N >=o Step 4 * C>=O Step 5 H2N N N C1N S) . 1 N NS) a H S)
75d Compound 75 75c Step 1: Ethyl (S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate (75a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5 g, 5.35 mmol) and potassium carbonate (1.4 g, 22.6 mmol) were dissolved in acetonitrile (20 mL), and (S) tetrahydrofuran-3-amine (660 mg, 5.35 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. Water was added and the organics were extracted three times with ethyl acetate, and was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)= 10:1) followed by concentration to obtain the title compound ethyl (S)-2-chloro-4 ((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate 75a (white solid, 2.2 g, 36% yield). 1H NMR (400 MHz DMSO) 6 8.68 (s, 1H), 8.58 (d, 1H), 4.79-4.77 (m, 1H), 4.39-4.33 (m, 2H), 4.04-3.98 (m, 2H), 3.90-3.87 (m, 1H), 3.76-3.73 (m, 1H), 2.43-2.33 (m, 1H), 1.95-1.88 (m, 1H), 1.39 (t, 3H). LC-MS m/z(ESI) = 272.00 [M+1]
Step 2: (S)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid (75b)
Ethyl (S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate 75a (2.2 g, 8.1 mmol) was dissolved in tetrahydrofuran/water (5 mL/5 mL), and lithium hydroxide (681 mg, 16.2 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v= 10/1) and concentrated to obtain the title compound (S)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid 75b (white solid, 1.7 g, 89% yield). 1H NMR (400 MHz DMSO) 6 13.83 (s, 1H), 8.65 (s, 1H), 8.63 (d, 1H), 4.61 (s, 1H), 3.89-3.80 (m, 2H), 3.76-3.70 (m, 1H), 3.65-3.63 (m, 1H), 2.50-2.25 (m, 1H), 1.87-1.86 (m, 1H). LC-MS m/z(ESI) = 244.20 [M+1]
Step 3: (S)-2-Chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (75c)
(S)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid 75b (1.7 g, 7 mmol) was dissolved in dimethylacetamide (10 mL), and triethylamine (707 mg, 7 mmol) and diphenylphosphoryl azide (1.9 g, 7 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1:6) to obtain the title compound (S)-2-chloro-9-(tetrahydrofuran-3-yl) 7,9-dihydro-8H-purin-8-one 75c (white solid, 1.4 g, 87% yield). 'H NMR (400 MHz DMSO) 6 8.13 (s, 1H), 5.00-4.93 (m, 1H), 4.12-4.06 (dd, 1H), 3.97 (t, 1H), 3.88-3.87 (m, 2H), 3.33 (s, 3H), 2.43-2.37 (m, 1H), 2.25-2.20 (m, 1H). LC-MS m/z(ESI)= 241.20 [M+1]
Step 4: (S)-2-Chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (75d)
(S)-2-Chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 75c (1.4 g, 5.8 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (735 mg, 5.8 mmol) and cesium carbonate (2.85 g, 8.7 mmol) were added at 0 °C. The reaction mixture was stirred for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to get the title compound (S)-2-chloro-7-methyl-9 (tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 75d (white solid, 1.2 g, 85% yield). 'H NMR (400 MHz DMSO) 6 8.35 (s, 1H), 5.04-4.97 (m, 1H), 4.13-4.07 (dd, 1H), 3.98 (t, 1H),3.90-3.84 (m, 2H), 3.35(s, 3H), 2.44-2.37 (m, 1H), 2.28-2.27 (m,1H). LC-MS m/z(ESI)= 255.20 [M+1]
Step 5: (S)-2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamiderile (compound 75)
(S)-2-Chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 57d (200 mg, 0.78 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (264 mg, 1.57 mmol), cesium carbonate (770 mg, 2.36 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (71.4 mg, 0.078 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound (S)-2-fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamiderile compound 75 (white solid, 58 mg, 19 % yield). 'H NMR (400 MHz DMSO) 6 8.49 (s, 1H), 8.20 (s, 1H), 7.89 (d, 1H),7.55 (d, 1H), 7.44(d, 2H), 4.95-5.02 (m, 1H), 4.07-4.13 (dd, 1H), 3.91-3.99(m,2H), 3.82 3.85(m,1H),3.32 (s, 3H), 2.44-2.50(m, 1H), 2.28(S, 3H), 2.20-2.27(m, 1 H). LC-MS m/z(ESI)= 387.10 [M+1]
Example 76
(R)-2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamiderile (compound 76)
0 H2 N - N N NAN N = F H
Compound 76 0 0 0 Step 1 N 0 '-6' Step 2 N OH Step 3 NII- O NH CI N O CIN NH C NH
1A 76a 76b
H 0 N N~ N c I o Step 4 > >O Step 5 H2N N N CI N N CIN NFa F "N N :N)
11 H 11
76d Compound 76 76c Step 1: Ethyl (R)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate (76a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5 g, 5.35 mmol) and potassium carbonate (1.4 g, 22.6 mmol) were dissolved in acetonitrile (20 mL), and (R) tetrahydrofuran-3-amine (660 mg, 5.35 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. Water was added and the organics were extracted three times with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)= 10:1) followed by concentration to obtain the title compound ethyl (R) 2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate 76a (white solid, 4.3 g, 65% yield). 'H NMR (400 MHz DMSO) 6 8.68 (s, 1H), 8.58 (d, 1H), 4.83-4.77 (m, 1H), 4.39-4.33 (m, 2H), 4.02-3.98 (m, 2H), 3.90-3.86 (m, 1H), 3.76-3.73 (m, 1H), 2.43-2.34 (m,1H), 1.98-1.88 (m, 1H), 1.39 (t, 3H). LC-MS m/z(ESI) = 272.00 [M+1]
Step 2: (R)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid (76b)
Ethyl (R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylate 76a (4.3 g, 15.8 mmol) was dissolved in tetrahydrofuran/water (5 mL/5 mL), and lithium hydroxide (1.3 g, 31.7 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v= 10/1) and concentrated to obtain the title compound
(R)-2-chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid 76b (white solid, 2.9 g, 87% yield). 'H NMR (400 MHz DMSO) 6 13.83 (s, 1H), 8.65 (s, 1H), 8.59 (d,1H), 4.62-4.60 (m, 1H), 3.89-3.80 (m, 2H), 3.76-3.70 (m, 1H), 3.64-3.56 (m, 1H), 2.33-2.23 (m, 1H), 1.88 1.83 (m, 1H). LC-MS m/z(ESI)= 244.20 [M+1]
Step 3: (R)-2-Chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (76c)
(R)-2-Chloro-4-((tetrahydrofuran-3-yl)amino)pyrimidine-5-carboxylic acid 76b (2.9 g, 11.9 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (1.2 g, 11.9 mmol) and diphenylphosphoryl azide (3.3 g, 11.9 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1:6) to obtain the title compound (R)-2-chloro-9 (tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 76c (white solid, 2.2 g, 68% yield). 'H NMR (400 MHz DMSO) 6 8.36 (s, 1H), 5.04-4.96 (m, 1H), 4.13-4.07 (dd, 1H), 3.97 (t, 1H), 3.87-3.84 (m, 2H), 3.33 (s, 3H), 2.40-2.36 (m, 1H), 2.29-2.27 (m, 1H). LC-MS m/z(ESI)= 241.20 [M+1]
Step 4: (R)-2-Chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (76d)
(R)-2-Chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 76c (2.2 g, 9.1 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (1.1 g, 9.1 mmol) and cesium carbonate (4.5 g, 13.7 mmol) were added at 0 °C. The reaction mixture was stirred for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to give the title compound (R)-2-chloro-7-methyl-9 (tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 76d (white solid, 1.7 g, 82% yield). 'H NMR (400 MHz DMSO) 6 8.41 (s, 1H), 5.05-4.97 (m, 1H), 4.13-4.08 (dd, 1H), 3.98 (t, 1H), 3.91-3.84 (m, 2H), 3.33 (s, 3H), 2.45-2.27 (m,1H), 2.26-2.20 (m,1H). LC-MS m/z(ESI)= 255.20 [M+1]
Step 5: (R)-2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamiderile (compound 76)
(R)-2-Chloro-7-methyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 76d (200 mg, 0.78 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (264 mg, 1.57 mmol), cesium carbonate (770 mg, 2.36 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (71.4 mg, 0.078 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound (R)-2-fluoro-5-methyl-4-((7-methyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H purin-2-yl)amino)benzamiderile compound 76 (white solid, 31 mg, 14.6 % yield). 1H NMR (400 MHz DMSO) 6 8.51 (s, 1H), 8.20 (s, 1H), 7.89 (d, 1H),7.54 (d, 1H), 7.44(d, 2H), 4.94-5.02 (m, 1H), 4.06-4.12 (dd, 1H), 3.87-3.98(m,2H), 3.82 3.85(m,1H),3.32 (s, 3H), 2.43-2.45(m, 1H), 2.33(s, 3H), 2.19-2.22(m, 1H). LC-MS m/z(ESI)= 387.10 [M+1]
Example 77 4-((9-(cis-4-Cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 77)
0 H2 N N N F N N N H
Compound 77 CN 0 OH 0 NH2 CN CN Step 1 Step 2 Step 3 Step 4
HN'Boc HN'Boc HN'Boc NH 2
77a 77b 77c 77d
0 OO H
N Step 5 , CI HOH Step 6 N C -=O Step 7 C1NI N NH - ~CI N N
CN CN CN 77e 77f 77g
0 N
CI O >= Step 8 H2N N N H
Compound 77 CN 77h N Step 1: tert-Butyl(cis-4-carbamoylcyclohexyl)carbamate (77b)
cis-4-((tert-Butoxycarbonyl)amino)cyclohexane-1-carboxylic acid 77a (5.0 g, 20.5 mmol) and O-(7-azabenzotriazolyl)-NN,N",V-tetramethyl-uronium hexafluorophosphate (9.4 g, 24.7 mmol) were dissolved in dichloromethane (15 mL). The solution was stirred at 0 °C for 20 min, and N,N-diisopropylethylamine (10.5 g, 82 mmol) and ammonium chloride (3.3 g, 61.5 mmol) were added. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, 10 mL of water was added to the reaction mixture, and the organic phase was separated, washed with saturated brine once, dried over anhydrous sodium sulfate and mixed with silica gel to prepare a sample, which was then purified by normal phase column chromatography to give a product, and the product was concentrated to obtain the title compound tert-butyl(cis-4 carbamoylcyclohexyl)carbamate 77b (white solid, 3.5 g, 71% yield). LC-MS m/z(ESI)= 243.30 [M+1]
Step 2: tert-Butyl(cis-4-cyanocyclohexyl)carbamate (77c)
tert-Butyl(cis-4-carbamoylcyclohexyl)carbamate 77b (3.5 g, 14.4 mmol) was dissolved in 70 mL of pyridine. The solution was cooled in an ice bath, and then phosphorus oxychloride (7.7 mL) was added dropwise. The reaction mixture was stirred in an ice bath for 1 h. The reaction was monitored by TLC. After completion of the reaction, 20 mL of water was added in an ice bath, and the resulting mixture was extracted 4 times with ethyl acetate. The organic phase was washed 7 times with acid water, finally washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to obtain the compound tert-butyl(cis-4 cyanocyclohexyl)carbamate 77c (yellow solid, 1.5 g, 36% yield). H NMR (400 MHz DMSO) 6 6.90 (d, 1H), 3.27-3.25 (m, 1H), 3.01-2.99 (m, 1H), 1.86 1.81 (m, 2H), 1.71-1.67 (m, 2H), 1.62-1.54 (m, 2H), 1.43-1.40 (m, 1H), 1.37(s, 3H),1.34-1.32(m, 1H). LC-MS m/z(ESI)= 225.30 [M+1]
Step 3: cis-4-Aminocyclohexane-1-carbonitrile (77d)
tert-Butyl(cis-4-cyanocyclohexyl)carbamate 77c (1.5 g, 6.7 mmol) was dissolved in hydrogen chloride in ethyl acetate (20 mL), and the reaction mixture was heated to °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, a white solid was precipitated. The reaction mixture was filtered and concentrated to dryness to give the title compound cis-4-aminocyclohexane-1 carbonitrile 77d (white solid, 1 g, 90% yield). LC-MS m/z(ESI)= 125.30 [M+1] Step 4: Ethyl 2-chloro-4-(cis-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate (77e)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (1.48 g, 6.7 mmol) and potassium carbonate (1.38 g, 10 mmol) were dissolved in acetonitrile (10 mL), and cis-4 aminocyclohexane-1-carbonitrile 77d (1 g, 6.7 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted three times with ethyl acetate, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)= 10:1) to obtain the title compound ethyl 2-chloro-4-(cis 4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate 77e (white solid, 589 mg, 60% yield). H NMR (400 MHz DMSO) 6 8.63 (s, 1H), 8.38 (d, 1H), 4.35-4.29 (m, 2H), 4.07-3.99 (m, 1H), 3.11-3.09 (m, 1H), 1.92-1.73 (m, 6H), 1,63-1.54 (m, 2H), 1.47(t, 3H). LC-MS m/z(ESI)= 310.20 [M+1]
Step 5:
2-Chloro-4-((cis-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid (77f)
Ethyl 2-chloro-4-((cis-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate 77e (589 mg, 1.9 mmol) was dissolved in tetrahydrofuran/water (4 mL/4 mL), and lithium hydroxide (160 mg, 3.8 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 2-chloro-4-((cis-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid 77f (white solid, 490 mg, 86% yield). 'H NMR (400 MHz DMSO) 6 13.83 (s, 1H), 8.62-8.60 (d, 2H), 4.05-3.98 (m, 1H), 3.10 (t, 1H), 1.94-1.73 (m, 6H), 1.60-1.56 (m, 2H). LC-MS m/z(ESI) = 282.30 [M+1]
Step 6: cis-4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (77g)
2-Chloro-4-((cis-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid 77f (490 mg, 1.75 mmol) was dissolved in dimethylacetamide (10 mL), and triethylamine (176 mg, 1.75 mmol) and diphenylphosphoryl azide (481 mg, 1.75 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:10) to obtain the title compound cis-4-(2 chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile 77g (white solid, 436 mg, 68% yield). H NMR (400 MHz DMSO) 6 11.64 (s, 1H), 8.13 (s, 1H), 4.23-4.16 (m, 1H), 3.22-3.16 (m, 1H), 2.44-2.39(m, 2H), 2.00 (d, 2H), 1.79-1.72 (m, 4H). LC-MS m/z(ESI) = 278.30 [M+1]
Step 7: cis-4-(2-Chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile (77h)
cis-4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile 77g (1.6 g, 5.77 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (728 mg, 5.77 mmol) and cesium carbonate (2.8 g, 8.65 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 30 min. The reaction was monitored by TLC. After completion of the reaction, 10 mL of water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to give the title compound cis-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile 77h (yellow solid, 1.25 g, 75% yield). H NMR (400 MHz DMSO) 6 8.35 (s, 1H), 4.28-4.19 (m, 1H), 3.34 (s, 3H), 3.23-3.17 (m, 1H), 2.46-2.39 (m, 2H), 2.00 (d, 2H),1.80-1.73 (m, 4H). LC-MS m/z(ESI)= 292.30 [M+1]
Step 8: 4-((9-(cis-4-Cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 77) cis-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile 77h (100 mg, 0.34 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (52 mg, 0.34 mmol), cesium carbonate (332 mg, 1.02 mmol) and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1' biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (31 mg, 0.034 mmol) were dissolved in dioxane (2 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(cis-4-cyanocyclohexyl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 77 (white solid, 44 mg, 29.3 % yield). 'H NMR (400 MHz DMSO) 6 8.36 (s, 1H), 8.17 (s, 1H), 7.89 (d, 1H), 7.52 (d, 1H), 7.44 (s, 1H), 7.26 (s, 1H), 4.20-4.26 (m, 1H), 3.32 (s, 3H), 3.19 (s, 1H), 2.44-2.47 (m, 2H), 2.26(s, 3 H), 2.02 (d, 2H), 1.67-1.81 (m, 4H). LC-MS m/z(ESI) = 424.20 [M+1]
Example 78 4-((9-(cis-4-cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-3 methylbenzonitrile (compound 78)
NC N N SNN N H
Compound 78 CN
N O NC NC N NO CI > N H -ONNN NZZZ 2 H
77h CN 5a Compound 78 CN cis-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile 77h (200 mg, 0.68 mmol), 4-amino-3-methylbenzonitrile 5a (90.7 mg, 0.68 mmol), cesium carbonate (664 mg, 2 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (62 mg, 0.068 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(cis-4-cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-3 methylbenzonitrile compound 78 (white solid, 120 mg, 60 % yield). 1H NMR (400 MHz DMSO) 6 8.48 (s, 1H), 8.22 (s, 1H), 8.20 (s, 1H),7.48 (d, 1H), 7.61
(s, 1H), 4.24-4.17 (m, 1H), 3.21 (s, 1H), 2.50-2.48 (m,2H), 2.32 (s,3H), 2.10-2.01 (m, 2H), 1.79-1.70 (m, 4H). LC-MS m/z(ESI) = 406.20 [M+1]
Example 79 4-((9-(trans-4-Cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 2-fluoro-5-methylbenzamide (compound 79)
0I
H2N HNF 2 ]::N NN N N~sz NO H
Compound 79 tN 0 OH 0 NH 2 CN CN Step 1 Step 2 Step 3 Step 4
H ,Boc H Boc H 'Boc NH 2
79a 79b 79c 79d
0 O H
CH Step 5 N HOH Step 6 C N Step 7
CI N NNH -J.- CI b H - CI N N
CN CN 79e 79f 79g
0 NN
>CO Step 8 H2N N H
tN Compound 79 tN 79h Step 1: tert-Butyl(trans-4-carbamoylcyclohexyl)carbamate (79b)
trans-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid 79a (5.0 g, 20.5 mmol) and O-(7-azabenzotriazolyl)-NN,N",V-tetramethyl-uronium hexafluorophosphate (9.4 g, 24.7 mmol) were dissolved in dichloromethane (15 mL). The solution was stirred at 0 °C for 20 min, and N,N-diisopropylethylamine (10.5 g, 82 mmol) and ammonium chloride (3.3 g, 61.5 mmol) were added. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, 10 mL of water was added to the reaction mixture, and the organic phase was separated, washed with saturated brine once, dried over anhydrous sodium sulfate and concentrated. The residue was purified by preparative medium pressure liquid chromatography to give the title compound tert-butyl(trans-4 carbamoylcyclohexyl)carbamate 79b (white solid, 4.4 g, 83% yield). LC-MS m/z(ESI) = 243.30 [M+1]
Step 2: tert-Butyl(trans-4-cyanocyclohexyl)carbamate (79c) trans-Butyl(cis-4-carbamoylcyclohexyl)carbamate 79b (4.4 g, 18.0 mmol) was dissolved in 70 mL of pyridine. The solution was cooled in an ice bath, and then phosphorus oxychloride (7.7 mL) was added to the reaction mixture dropwise. The reaction mixture was stirred in an ice bath for 1 h. The reaction was monitored by TLC. After completion of the reaction, 20 mL of water was added in an ice bath, and the resulting mixture was extracted 4 times with ethyl acetate. The organic phase was washed 7 times with acid water, finally washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the compound tert butyl(trans-4-cyanocyclohexyl)carbamate 79c (yellow solid, 1.2 g, 30% yield). H NMR (400 MHz DMSO) 6 6.80 (m, 1H), 3.24-3.22 (m, 1H), 2.63-2.55 (m, 1H), 1.99-1.95 (m, 2H),1.77-1.73 (m, 2H), 1.56-1.46 (m, 2H), 1.36 (s, 9H), 1.21-1.11 (m, 2H). LC-MS m/z(ESI)= 225.30 [M+1]
Step 3: trans-4-Aminocyclohexane-1-carbonitrile (79d)
tert-Butyl(cis-4-cyanocyclohexyl)carbamate 79c (1.2 g, 9.6 mmol) was dissolved in hydrogen chloride in ethyl acetate (20 mL), and the reaction mixture was heated to °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, a white solid was precipitated. The reaction mixture was filtered and concentrated to obtain the title compound trans-4-aminocyclohexane-1-carbonitrile 79d (white solid, 660 mg, 60% yield). LC-MS m/z(ESI)= 125.30 [M+1]
Step 4: Ethyl 2-chloro-4-(trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate (79e)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (1.4 g, 5.35 mmol) and potassium carbonate (1.4 g, 10.7 mmol) were dissolved in acetonitrile (10 mL), and trans-4 aminocyclohexane-1-carbonitrile 79d (660 mg, 5.35 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted three times with ethyl acetate, and the organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)= 10:1) to obtain the title compound ethyl 2-chloro-4-(trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate 79e (white solid, 810 mg, 62% yield). 'H NMR (400 MHz DMSO) 6 8.62 (s, 1H), 8.27 (d, 1H), 4.30 (q, 2H), 4.04-3.96 (m, 1H), 4.04-3.67 (m, 1H), 2.76-2.70 (m, 1H), 2.04-2.00 (m, 2H), 1.96-1.92 (m, 2H), 1.72 1.62 (m, 2H), 1.47-1.37(m, 2H), 1.30(t,3H). LC-MS m/z(ESI)= 310.20 [M+1]
Step 5: 2-Chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid (79f)
Ethyl 2-chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylate 79e (810 mg, 2.6 mmol) was dissolved in tetrahydrofuran/water (4 mL/4 mL), and lithium hydroxide (247 mg, 5.2 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitate. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 2-chloro-4-((trans 4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid 79f (white solid, 790 mg, 86% yield). H NMR (400 MHz DMSO) 6 8.58 (s, 1H), 8.49 (d, 1H), 4.01-3.87 (m,1H), 2.76-2.71 (m, 1H), 2.03-1.93 (m, 4H), 1.72-1.63 (m, 2H), 1.44-1.35 (m, 2H). LC-MS m/z(ESI) = 282.30 [M+1]
Step 6: trans-4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile (79g)
2-Chloro-4-((trans-4-cyanocyclohexyl)amino)pyrimidine-5-carboxylic acid 79f (730 mg, 2.6 mmol) was dissolved in dimethylacetamide (10 mL), and triethylamine (263 mg, 2.6 mmol) and diphenylphosphoryl azide (715 mg, 2.6 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:10) to obtain the title compound trans-4-(2 chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile 79g (white solid, 553 mg, 68% yield). H NMR (400 MHz DMSO) 6 11.63 (s, 1H), 8.12 (s, 1H), 4.23-4.16 (m, 1H), 2.78-2.72 (m, 1H), 2.25-2.12 (m, 4H), 1.82-1.68 (m, 4H). LC-MS m/z(ESI) = 278.30 [M+1]
Step 7: trans-4-(2-Chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile (79h)
trans-4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1-carbonitrile 79g (553 mg, 2 mmol) was dissolved in dimethylformamide (5 mL), and dimethyl sulfate (252 mg, 2 mmol) and cesium carbonate (977 mg, 3 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 30 min. The reaction was monitored by TLC. After completion of the reaction, 10 mL of water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration to give the title compound trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane 1-carbonitrile 79h (yellow solid, 420 mg, 72% yield). H NMR (400 MHz DMSO) 6 8.34 (s, 1H), 4.28-4.21 (m, 1H), 2.79-2.72 (m, 1H), 2.22 2.13 (m, 4H), 1.82-1.70 (m, 4H). LC-MS m/z(ESI)= 292.30 [M+1]
Step 7: 4-((9-(trans-4-Cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 2-fluoro-5-methylbenzamide (compound 79)
trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile 79h (100 mg, 0.34 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (52 mg, 0.34 mmol), cesium carbonate (332 mg, 1.02 mmol) and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1' biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (31 mg, 0.034 mmol) were dissolved in dioxane (2 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(trans-4-cyanocyclohexyl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 79 (white solid, 57 mg, 36.2% yield). 1H NMR (400 MHz DMSO) 6 8.48 (s, 1H), 8.18 (s, 1H), 7.95 (d, 1H), 7.54 (d, 1H), 7.45 (d, 1H), 4.20-4.29 (m, 1H), 3.31 (s, 3H), 2.60-2.67 (m, 1H), 2.25-2.34 (m, 5H), 2.15(d, 2 H), 1.83 (d, 2H), 1.72-1.76 (m, 2H). LC-MS m/z(ESI) = 424.20 [M+1]
Example 80 4-((9-(trans-4-cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 3-methylbenzonitrile (compound 80)
NC N NO -NN N HO
Compound 80 CN
N NC N N CK'D N I NCa O X "
NH2 N N H NV
7N Sa Compound 80 CN 79h5at trans-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexane-1 carbonitrile 79h (200 mg, 0.68 mmol), 4-amino-3-methylbenzonitrile 5a (90.7 mg, 0.68 mmol), cesium carbonate (664 mg, 2 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (62 mg, 0.068 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(trans-4-cyanocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) 3-methylbenzonitrile compound 80 (white solid, 120 mg, 60% yield). 1H NMR (400 MHz DMSO) 6 8.60 (s, 1H), 8.16 (s, 1H), 8.12 (d, 1H),7.68 (d, 1H), 7.62
(s, 1H), 4.26-4.18 (m, 1H), 3.31 (s, 3H), 2.67-2.60 (m,1H), 2.32 (s,3H), 2.31-2.27(m, 2H), 2.24-2.07(m, 2H), 1.76 (dd, 2H), 1.75-1.64 (m, 2H). LC-MS m/z(ESI)= 406.20 [M+1]
Example 81 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(piperidin-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide (compound 81)
H 2N N N H
Compound 81 H
0 0 H
N ON- O N OH N NO Step CI N NH Step 2 CI N NH Step 3 CI N N Cl, 1CI N CI NH N
N N N N Boo Boc Boc IA 81a 81b 81c
I 0 I0 NN Stept4 eIp H2 N N5 = Step 6 H2N N O F NNN N1 N== F H Nb H N N Boo 81d 81e Boc Compound 81 H
Step 1: Ethyl 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5 carboxylate (81a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5 g, 22.6 mmol) and potassium carbonate (6.2 g, 45.2 mmol) were dissolved in acetonitrile (20 mL), and tert-butyl 4 aminopiperidine-1-carboxylate (4.5 g, 22.6 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted three times with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v)= 10:1) followed by concentration to obtain the title compound ethyl 4-((1-(tert butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylate 81a (white solid, 8.2 g, 95% yield). 1H NMR (400 MHz DMSO) 6 8.63 (s, 1H), 8.32 (d, 1H), 4.33-4.28 (m, 2H), 4.17-4.14 (m, 1H), 3.85 (d, 2H), 2.94 (s, 2H), 1.88-1.80 (m, 2H), 1.51-1.44 (m, 2H), 1.41 (s, 9H), 1.32-1.29 (m, 3H). LC-MS m/z(ESI) = 385.10 [M+1]
Step 2: 4-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid (81b)
Ethyl 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5 carboxylate 81a (8.2 g, 21.3 mmol) was dissolved in tetrahydrofuran/water (10 mL/5 mL), and lithium hydroxide (1.8 g, 42.7 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 4-((1-(tert butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid 81b (white solid, 7 g, 86% yield). 'H NMR (400 MHz DMSO) 6 8.59 (s, 1H), 8.54 (d, 1H), 4.20-4.10 (m,1H), 3.87-3.84 (m, 2H), 2.96 (s, 3H), 1.91-1.73 (m, 3H), 1.41(s, 9H). LC-MS m/z(ESI)= 357.10 [M+1]
Step 3: tert-Butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate (81c)
4-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid 81b (7 g, 19.6 mmol) was dissolved in dimethylacetamide (10 mL), and triethylamine (1.96 g, 19.6 mmol) and diphenylphosphoryl azide (5.4 g, 19.6 mmol) were added. The reaction mixture was gradually heated to 110 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:10) to obtain the title compound tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1 carboxylate 81c (white solid, 6.4 g, 87% yield). 'H NMR (400 MHz DMSO) 6 8.14 (s, 1H), 4.41-4.33 (m, 1H), 4.06 (d, 2H), 2.88 (s, 2H), 2.30-2.20 (m, 2H), 1.84-1.71 (m, 2H), 1.43 (s, 9H). LC-MS m/z(ESI)= 354.10 [M+1]
Step 4: tert-Butyl 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1 carboxylate (81d)
tert-Butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate 81c (6.4 g, 18.1 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (2.28 g, 18.1 mmol) and cesium carbonate (8.5 g, 27.1 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The mixture was filtered to give the title compound tert-butyl 4-(2 chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate 81d (white solid, 5.4 g, 79% yield). 'H NMR (400 MHz DMSO) 6 8.36 (s, 1H), 4.46-4.37 (m, 1H), 4.05 (d, 2H), 3.35 (s, 3H), 2.87 (s, 2H), 2.33-2.19 (m, 2H), 1.74-1.72 (m, 2H), 1.43 (s, 9H). LC-MS m/z(ESI) = 368.10 [M+1]
Step 5: tert-Butyl 4-(2-((4-carbamoyl-5-fluoro-2-methylphenyl)amino)-7-methyl-8-oxo-7,8 dihydro-9H-purin-9-yl)piperidine-1-carboxylate (81e)
tert-Butyl 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1 carboxylate 81d (200 mg, 0.54 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (181 mg, 0.54 mmol), cesium carbonate (528 mg, 1.08 mmol) and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1' biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (50 mg, 0.054 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 60/1) to obtain the title compound tert-butyl 4-(2-((4-carbamoyl-5-fluoro-2 methylphenyl)amino)-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1 carboxylate 81e (white solid, 119 mg, 58% yield). 1H NMR (400 MHz DMSO) 6 8.52 (s, 1H), 8.18 (s, 1H), 7.85 (d, 1H), 7.53 (d, 1H), 7.31 (s, 1H), 4.40-4.34 (m, 1H), 4.10 (dd, 2H), 3.20 (s, 3H), 2.88-2.74 (m, 2H), 2.37 2.30 (m, 2H), 2.27 (s, 3H), 1.75 (d, 2H), 1.40 (s, 9 H). LC-MS m/z(ESI)= 500.20 [M+1]
Step 6: 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(piperidin-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide (compound 81)
tert-Butyl 4-(2-((4-carbamoyl-5-fluoro-2-methylphenyl)amino)-7-methyl-8-oxo-7,8 dihydro-9H-purin-9-yl)piperidine--carboxylate 81e (119 mg, 0.23 mmol) was added to a 50 mL single-necked flask, and ethyl acetate hydrochloride 2 M (10 mL) was added thereto. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by Pre-HPLC to obtain the title compound 2-fluoro-5-methyl-4-((7-methyl-8-oxo-9-(piperidin-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide compound 81 (white solid, 32 mg, 23% yield). 1H NMR (400 MHz DMSO) 6 8.48 (s, 1H), 8.17 (d, 1H), 7.94 (d, 1H), 7.54 (d, 1H), 7.45-7.37 (m, 1H), 4.29-4.21 (m, 1H), 3.32 (s, 3H), 3.08-3.05 (m, 2H), 2.57-2.51 (m, 2H), 3.39-3.32 (m, 2H), 2.29 (s, 3H), 1.68-1.64 (m, 2H). LC-MS m/z(ESI) = 400.20 [M+1]
Example 82 2-Fluoro-5-methyl-4-((7-methyl-9-(1-methylpiperidin-4-yl)-8-oxo-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 82)
0
H2N N'N N H
Compound 82
N N 0
CI Step 1 I Step 3 H2N N Sp 1 N F N N N H b N N Boc HN 81d H2 2 Compound 82 b
Step 1: 2-Chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8 118>=o -one (82a) 'Dc)= tert-Butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidine-1-carboxylate 81d (2 g,5.4inmol) was added to areaction flask, and 2 Mhydrochloric acid-ethyl acetate solution (10 mL) was added with stirring at room temperature. The reaction mixture was stirred for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to give the hydrochloride salt of the title compound 2-chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one as hydrochloride salt 82a (white solid, 1.4 g, 91% yield). 'H NMR (400 MHz DMSO) 6 8.39 (s, 1H), 6.42 (s, 1H), 4.59-4.53 (m, 1H), 3.39 (s, 2H), 3.36 (s, 3H), 3.16-3.04 (m, 2H), 2.62- 2.50 (m, 2H), 2.07-1.93 (m, 2H). LC-MS m/z(ESI) = 268.10 [M+1]
Step 2: 2-Chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one (82b)
2-Chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one 82a (1.4 g, 5.4 mmol) was dissolved in methanol (20 mL), and 4A molecular sieve (100 mg) was added, followed by the addition of polyformaldehyde (783 mg, 27 mmol). The reaction mixture was stirred at room temperature for 6 h. Sodium cyanoborohydride (1 g, 16.2 mmol) was added, and the reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated to give the title compound 2-chloro-7-methyl-9-(1 methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one 82b (white solid, 600 mg, 62% yield). H NMR (400 MHz DMSO) 6 8.35 (s, 1H), 4.21-4.13 (m, 1H), 3.35 (s, 3H), 2.92 (d, 2H), 2.45-2.41 (m, 2H), 2.23 (s, 3H), 2.09-2.03 (m, 2H), 1.70-1.67 (m, 2H). LC-MS m/z(ESI) = 282.10 [M+1]
Step 3: 2-Fluoro-5-methyl-4-((7-methyl-9-(1-methylpiperidin-4-yl)-8-oxo-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 82)
2-Chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one 82b (150 mg, 0.53 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (178 mg, 1.06 mmol), cesium carbonate (518 mg, 1.59 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (48 mg, 0.053 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1) followed by Pre-HPLC to obtain the title compound 2-fluoro-5-methyl-4-((7-methyl-9-(1 methylpiperidin-4-yl)-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 82 (white solid, 20 mg, 18% yield). 'H NMR (400 MHz DMSO) 6 8.27 (s, 1H), 8.14 (s, 1H), 7.93-7.89 (d, 1H), 7.58-7.55 (d, 1H),7.22 (s,2H), 3.34 (s, 3H), 2.95-2.92 (d, 2H), 2.67-2.55 (m, 2H), 2.30 (s, 3H), 2.25 (s, 3H), 2.10-2.04 (m, 2H), 1.91 (s, 1H), 1.72-1.68 (d, 2H). LC-MS m/z(ESI) = 414.20 [M+1]
Example 83 4-((9-(4,4-Difluorocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 83)
H2 N N F N NN H
Compound 83 F F 0 0 0
N O N O N OH Step CI N NH Step 2 C N NH Step 3 CI -N CI ~ C1 N NH
F F F F
IA 83a 83b
0 H N F 0 NI>= N Se = H 2N N N==N NN
N FI F F
83c F 83d Compound 83 F
Step 1: Ethyl 2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylate (83a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (6.5 g, 29.41 mmol) and 4,4 difluorocyclohexa-1-amine hydrochloride (5.0 g, 29.41 mmol) were dissolved in acetonitrile (100 mL), and potassium carbonate (10.16 g, 73.52 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filter residue was washed with ethyl acetate. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v) = 1:1) to obtain the title compound ethyl 2-chloro-4-((tetrahydro-2H-pyran-4 yl)amino)pyrimidine-5-carboxylate 83a (white solid, 8.0 g, 85.10% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.85 (s, 1H), 8.11 (d, 1H), 4.34-4.25 (in, 2H), 4.21
4.10 (in, 1H), 2.11-1.91 (in, 6H), 1.71-1.57 (in, 2H), 1.23 (t, 3H). LC-MS m/z (ESI) = 320.10 [M+l]
Step 2: 2-Chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylic acid (83b)
Ethyl 2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylate 83a (4 g, 12.47 mmol) was dissolved in tetrahydrofuran/water (50 mL/50 mL), and lithium hydroxide (597 mg, 24.94 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 5 with 2 N hydrochloric acid, and a while solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to give the title compound 2-chloro-4-((4,4-difluorocyclohexyl)amino)pyrimidine-5-carboxylic acid 83b (white solid, 3.4 g, 91.53% yield).
H NMR (400 MHz, DMSO-d6) 6 13.79 (s, 1H), 8.60 (s, 1H), 8.55 (d, 1H), 4.22-4.06 (m, 1H), 2.12-1.89 (m, 6H), 1.72-1.56 (m, 2H). LC-MS m/z (ESI) = 292.00 [M+1]
Step 3: 2-Chloro-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one (83c)
2-Chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid 83b (3.34 g, 11.45 mmol) was dissolved in dimethylacetamide (50 mL), and triethylamine (1.6 mL, 11.45 mmol) and diphenylphosphoryl azide (1.22 mL, 11.45 mmol) were added. The reaction mixture was gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. The reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, and concentration and drying were performed in vacuo to give the title compound 2-chloro-9-(4,4 difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one 83c (white solid, 2.2 g, 78.5% yield). H NMR (400 MHz, DMSO-d6) 6 11.66 (s, 1H), 8.13 (s, 1H), 4.46-4.36 (m, 1H), 2.17 1.94 (m, 6H), 1.87-1.80 (m, 2H). LC-MS m/z (ESI) = 289.10 [M+1]
Step 4: 2-Chloro-9-(4,4-difluorocyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (83d)
2-Chloro-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one 83c (2.62 g, 9.08 mmol) was dissolved in dimethylformamide (50 mL), and dimethyl sulfate (1.14 g, 9.08 mmol) and cesium carbonate (4.44 g, 13.61 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to yield the title compound 2 chloro-7-methyl-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one 83d (white solid, 2.2 g, 80% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.37 (s, 1H), 4.55-4.39 (m, 1H), 3.33 (s, 3H), 2.21 2.01 (m, 6H), 1.90-1.79 (m, 2H). LC-MS m/z (ESI) = 303.10 [M+1]
Step 5: 4-((9-(4,4-Difluorocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 83)
2-Chloro-7-methyl-9-(4,4-difluorocyclohexyl)-7,9-dihydro-8H-purin-8-one 83d (150 mg, 0.495 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (166.6 mg, 0.991 mmol), cesium carbonate (323 mg, 0.991 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (45 mg, 0.05 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 4-((9-(4,4 difluorocyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5 methylbenzamide compound 83 (white solid, 30 mg, 14% yield).
H NMR (400 MHz, DMSO-d6) 6 8.49 (s, 1H), 8.20 (s, 1H), 7.84 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.38-7.33 (m, 1H), 4.45-4.38 (m, 1H), 3.32 (s, 3H), 2.28 (s, 3H), 2.18-1.94 (m, 6H), 1.87-1.81 (m, 2H). LC-MS m/z (ESI) = 435.10 [M+l]
Example 84 2-Fluoro-4-((9-((3aR,6aS)-5-hydroxyoctahydropentalen-2-yl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide(compound84)
0 H 2N N NO F ) NN N H H H
OH
Compound 84 0 0 / OH 0O N/ O N
H Step H Step 2 H Step 3 ,C >N Step 4 CI N N Step 5 H H H H H H H H O NH 2 O O
84a 84b 84c 84d 84e
COCIN OCIN N OI2 N Step 6 Ste 7 Step 8 F H H H H H HH H
O\O O OH OH
84f 84g 84h Compound 84
Step 1: (3aR,6aS)-Tetrahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5(3H)-one (84b)
(3as,6as)-Tetrahydropyrene-2,5(1H,3H)-dione 84a (20 g, 145 mmol) and p toluenesulfonic acid (2.7 g, 14.5 mmol) were added to a dry reaction flask and dissolved in toluene (250 mL), and ethylene glycol (7.3 mL, 130 mmol) was added dropwise at 120 °C. After addition, the reaction mixture was refluxed for 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 4/1) to obtain the title compound (3aR,6aS) -tetrahydro-H-spiro[pentalene-2,2'
[1,3]dioxolan]-5(3H)-one 84b (pale yellow liquid, 10.8 g, 40.88% yield). LC-MS m/z (ESI) = 183.10 [M+1]
Step 2: (3aR,6aS)-Hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5-amine (84c)
(3aR,6aS)-Tetrahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5(3H)-one 84b (10.8 g, 59 mmol) was dissolved in methanol (100 mL), and ammonium acetate (45 g, 590 mmol) was added. The reaction mixture was stirred at 55 °C for 2 h. Sodium cyanoborohydride (5.5 g, 88.5 mmol) was added, and the reaction mixture was stirred for another 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 10/1) to obtain the title compound (3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5-amine 84c (crude, yellow liquid, 5.5 g, 17.82% yield). LC-MS m/z (ESI) = 184.10 [M+1]
Step 3: Ethyl 2-chloro-4-(((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5 yl)amino)pyrimidine-5-carboxylate (84d)
(3aR,6aS)-Hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolane]-5-amine 84c (5.5 g, 30.03 mmol) and potassium carbonate (12.4 g, 90.1 mmol) were well mixed with acetonitrile (100 mL), and ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (9.9 g, 45.05 mmol) was added at 0 °C. The reaction mixture was warmed to room temperature and stirred overnight. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered. The filtrate was collected and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 8/1) to obtain the title compound ethyl 2-chloro-4-(((3aR,6aS) hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxino]-5-yl)amino)pyrimidine-5 carboxylate 84d (pale yellow solid, 0.74 g, 6.70% yield). LC-MS m/z (ESI) = 368.10 [M+1]
Step 4: 2-Chloro-4-(((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5 yl)amino)pyrimidine-5-carboxylic acid (84e)
Ethyl 2-chloro-4-(((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxan]-5 yl)amino)pyrimidine-5-carboxylate 84d (0.74 g, 2.01 mmol) was dissolved in tetrahydrofuran/water (1/1, 40 mL), and lithium hydroxide monohydrate (0.24 g, 6.03 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 3 with 2 N hydrochloric acid, and a white solid was precipitated. The mixture was filtered, and the filter cake was collected and dried to yield the title compound 2-chloro-4-(((3aR,6aS)-hexahydro-1H spiro[pentalene-2,2'-[1,3]dioxido]-5-yl)amino)pyrimidine-5-carboxylic acid 84e (white solid, 0.5 g, 73.20% yield). LC-MS m/z (ESI) = 340.10 [M+1]
Step 5: 2-Chloro-9-((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5-yl)-7,9 dihydro-8H-purin-8-one (84f)
2-Chloro-4-(((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxido]-5 yl)amino)pyrimidine-5-carboxylicacid 84e (0.5 g, 1.47 mmol) and diphenylphosphoryl azide (0.41 g, 1.47 mmol) were added to a dry reaction flask and dissolved in N,N dimethylacetamide (20 mL), and triethylamine (0.2 mL, 1.47 mmol) was added dropwise at 0 °C. After addition, the reaction mixture was warmed to room temperature and stirred for 30 min, and then stirred at 120 °C for 2 h. The reaction was monitored by TLC. The reaction mixture was poured into six volumes of ice water, and a pale yellow solid was precipitated. The mixture was filtered, and the filter cake was washed with water (60 mL) and dried to give the target compound 2-chloro-9-((3aR,6aS) hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxino]-5-yl)-7,9-dihydro-8H-purin-8-one 84f (pale yellow solid, 0.38 g, 76.77% yield). LC-MS m/z (ESI) = 337.10 [M+1]
Step 6: 2-Chloro-9-((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxolan]-5-yl)-7 methyl-7,9-dihydro-8H-purin-8-one (84g)
2-Chloro-9-((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxido]-5-yl)-7,9 dihydro-8H-purin-8-one 84f (0.38 g, 1.13 mmol), dimethyl sulfate (0.14 g, 1.13 mmol) and cesium carbonate (0.55 g, 1.69 mmol) were added to a dry reaction flask and dissolved in N,N-dimethylformamide (20 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic phases were combined, dried and concentrated to yield the title compound 2 chloro-9-((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxino]-5-yl)-7-methyl 7,9-dihydro-8H-purin-8-one 84g (yellow liquid, crude, 0.68 g, 174% yield). LC-MS m/z (ESI) = 351.10 [M+1]
Step 7: 2-Chloro-9-((3aR,6aS)-5-hydroxyoctahydropentalen-2-yl)-7-methyl-7,9-dihydro-8H purin-8-one (84h)
2-Chloro-9-((3aR,6aS)-hexahydro-1H-spiro[pentalene-2,2'-[1,3]dioxido]-5-yl)-7 methyl-7,9-dihydro-8H-purin-8-one 84g (0.39 g, 1.13 mmol, theoretical) are dissolved in tetrahydrofuran/methanol (10/1, 11 mL), and diluted hydrochloric acid (3.8 mL, 3 mol/L) was added. The reaction mixture was stirred at room temperature for 1.5 h, concentrated to remove tetrahydrofuran and methanol, the pH was adjusted to about 8 with saturated sodium bicarbonate, and extracted with ethyl acetate. The organic phases were combined, dried and concentrated to yield a yellow solid (0.37 g), which was then dissolved in tetrahydrofuran/methanol (10/1, 11 mL), and sodium borohydride (0.05 g, 1.35 mmol) was added. The mixture was obtain at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction system was quenched with water (0.5 mL) and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1/2) to obtain the title compound 2-chloro-9 ((3aR,6aS)-5-hydroxyoctahydropentalen-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 84h (white solid, 0.27 g, 77.38% yield).
Step 8: 2-Fluoro-4-((9-((3aR,6aS)-5-hydroxyoctahydropentalen-2-yl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide (compound 84)
2-Chloro-9-((3aR,6aS)-5-hydroxyoctahydropenten-2-yl)-7-methyl-7,9-dihydro-8H purin-8-one 84h (0.19 g, 0.61 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (0.41 g, 2.46 mmol), cesium carbonate (0.8 g, 2.46 mmol) and Brettphos G3 Pd (56 mg, 0.061 mmol) were dissolved in 1,4-dioxane (20 mL), and the system was purged with nitrogen three times. The reaction mixture was stirred at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (DCM/MeOH (v/v) = 30/1) to obtain the title compound 2-fluoro-4-((9-((3aR,6aS)-5 hydroxyoctahydropentan-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) -methylbenzamide compound 84 (white solid, 0.14 g, 52.10% yield). 1H NMR (600 MHz, DMSO-d) 68.49 (s, 1H), 8.17 (s, 1H), 7.82 (d, 1H), 7.54 (d, 1H), 7.48 (s, 1H), 7.37 (s, 1H), 4.74 (s, 1H), 4.61-4.52 (m, 1H), 4.09-3.99 (m, 1H), 3.31 (s, 3H), 2.36-2.30 (m, 4H), 2.07-1.98 (m, 4H), 1.37-1.28 (m, 3H), 1.23 (d, 2H). LC-MS m/z (ESI) = 441.20 [M+1]
Example 85 4-((9-(8-Oxabicyclo[3.2.1]octan-3-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 85)
0 H2 N N N F N' N N>= HO
Compound 85 0 0
Step 1 NH Step 2 Step 3 N H0 Step4 N HOH 0 N0~ NH2 CI N' NH CI N- NH
85a 85b 85c 85d 85e
H 0
Step 5 :sCIO Step 6>:CIO Step 7 H 2N N N cr'N" N nCI N N = F N N N>= H 6 85f 85g Compound 85
Step 1: 8-Oxabicyclo[3.2.1]octan-3-one oxime (85b)
8-Oxabicyclo[3.2.1]octan-3-one 85a (1.4 g, 11.1 mmol), hydroxylamine hydrochloride (925 mg, 13.3 mmol) and potassium carbonate (3.1 g, 22.2 mmol) were dissolved in a solvent mixture of ethanol/water (10/5 mL). The reaction mixture was stirred at 80 °C for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove ethanol, and water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated to give the title compound 8 oxabicyclo[3.2.1]octan-3-oneoxime85b (pale yellow solid, crude, 1.56g, 100% yield). LC-MS m/z(ESI) = 142.10 [M+1]
Step 2: 8-Oxabicyclo[3.2.1]octan-3-amine (85c)
8-Oxabicyclo[3.2.1]octan-3-one oxime 85b (1.5 g, 10.64 mmol) was dissolved in methanol (30 mL), and nickel chloride hexahydrate (2.53 g, 10.64 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 0.5 h and then cooled to -30 °C. Sodium borohydride (6.0 g, 159.6 mmol) was slowly added, and after addition, the reaction mixture was slowly warmed to room temperature and stirred overnight. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate, and the organic layer was concentrated to yield the title compound 8-oxabicyclo[3.2.1]octan-3-amine 85c (pale yellow oil, 639 mg, 45% yield). LC-MS m/z(ESI) = 128.10 [M+1]
Step 3: Ethyl 4-((8-oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylate (85d)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (1.1 g, 5.03 mmol) and potassium carbonate (1.74 g, 12.58 mmol) were dissolved in acetonitrile (20 mL), and 8 oxabicyclo[3.2.1]octan-3-amine 85c (639 mg, 5.03 mmol) was added at 0 °C. The reaction mixture was stirred at room temperature for 20 h. The reaction was monitored by TLC. After completion of the reaction, water (30 mL) was added to the reaction mixture, and a solid was precipitated. The mixture was filtered, and the filter cake was washed 3 times with water and dried to give the title compound ethyl 4-((8 oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylate 85d (white solid, 1.01 g, 64.3% yield). 'H NMR (400 MHz, DMSO) 6 8.91 (d, 1H), 8.64 (s, 1H), 4.38-4.31 (m, 4H), 4.30-4.24 (m, 1H), 2.20-2.08 (m, 2H), 2.02-1.90 (m, 4H), 1.66 (d, 2H), 1.32 (t, 3H). LC-MS m/z(ESI) = 312.10 [M+1]
Step 4: 4-((8-Oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid (85e)
Ethyl 4-((8-oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylate d (1.0 g, 3.21 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL of water, and lithium hydroxide (308 mg, 12.83 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 4, and a white solid precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1), concentrated and dried to obtain the title compound 4-((8-oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5 carboxylic acid 85e (white solid, 808 mg, 87.9% yield). 'H NMR (400 MHz, DMSO) 6 13.83 (s, 1H), 9.17 (d, 1H), 8.59 (s, 1H), 4.32 (s, 2H), 4.29-4.22 (m, 1H), 2.17-2.07 (m, 2H), 2.00-1.89 (m, 4H), 1.65 (d, 2H). LC-MS m/z(ESI)= 284.20 [M+1]
Step 5: 9-(8-Oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one (85f)
4-((8-Oxabicyclo[3.2.1]octan-3-yl)amino)-2-chloropyrimidine-5-carboxylic acid 85e (808 mg, 2.85 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (288 mg, 2.85 mmol) and diphenylphosphoryl azide (784 mg, 2.85 mmol) were added. The reaction mixture was gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 5:1 to 1:10) to obtain the title compound 9-(8-oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7,9 dihydro-8H-purin-8-one 85f (white solid, 653 mg, 71% yield). 'H NMR (600 MHz, DMSO) 6 11.64 (s, 1H), 8.12 (s, 1H), 4.47-4.44 (m, 2H), 4.42 4.40 (m, 1H), 2.28-2.20 (m, 2H), 2.07-2.01 (m, 2H), 1.96-1.90 (m, 2H), 1.81-1.75 (m, 2H). LC-MS m/z(ESI)= 281.10 [M+1]
Step 6: 9-(8-Oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one (85g)
9-(8-Oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7,9-dihydro-8H-purin-8-one 85f (653 mg, 2.33 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (293 mg, 2.33 mmol) and cesium carbonate (1.52 g, 4.65 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, 30 mL of water and ethyl acetate was added to the reaction mixture, and the organic phase was concentrated to afford the title compound 9-(8 oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one 85g (white solid, 533 mg, 81.6% yield). 'H NMR (400 MHz, DMSO) 6 8.36 8.34 (m, 1H), 4.51-4.47 (m, 2H), 4.45-4.42 (m, 1H), 3.34 (s, 3H), 2.31-2.22 (m, 2H), 2.07-1.98 (m, 2H), 1.98-1.90 (m, 2H), 1.79 (t, 2H). LC-MS m/z(ESI)= 295.20 [M+1]
Step 7: 4-((9-(8-Oxabicyclo[3.2.1]octan-3-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 85)
9-(8-Oxabicyclo[3.2.1]octan-3-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one g (100 mg, 0.34 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (57 mg, 0.34 mmol), cesium carbonate (221 mg, 0.68 mmol) and Brettphos Pd G3 (31 mg, 0.034 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 4-((9-(8 oxabicyclo[3.2.1]octan-3-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide compound 85 (white solid, 41 mg, 28.9% yield). 'H NMR (400 MHz, DMSO) 6 8.79 (s, 1H), 8.24-8.16 (m, 1H), 7.75 (d, 1H), 7.57 (d, 1H), 7.50 (s, 1H), 7.42 (s, 1H), 4.53-4.37 (m, 3H), 2.27 (s, 3H), 2.24-2.15 (m, 2H), 2.03
(t, 2H), 1.87 (s, 2H), 1.67 (d, 2H). LC-MS m/z(ESI) = 427.20 [M+1]
Example 86 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(2-oxaspiro[3.5]nonan-7-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 86)
0/ H2N ' N~ N>= FH2 N N H
0 Compound 86 0 0 HO'N NH2 NN O N OH
Step I Step 2 Step 3 CI N NH Step 4 CI N NH
0 0 0 0
86a 86b 86c 86d 86e
H 0
Step 5 Step 6 I O Step 7 H2N N N O C N N -CINN I-N N N>= H
0 0 0 86f 86g Compound 86 Step 1: 2-Oxaspiro[3.5]nonan-7-one oxime (86b)
2-Oxaspiro [3.5]nonan-7-one 86a (1.5 g, 10.7 mmol), hydroxylamine hydrochloride (744 mg, 10.7 mmol) and potassium carbonate (2.95 g, 21.4 mmol) were dissolved in a solvent mixture of ethanol/water (10/5 mL). The reaction mixture was stirred at 80 °C for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated to obtain the title compound 2-oxaspiro[3.5]nonan-7-one oxime 86b (pale yellow solid, crude, 1.45 g, 89% yield). LC-MS m/z(ESI)= 156.10 [M+1]
Step 2: 2-Oxaspiro[3.5]nonan-7-amine (86c)
2-Oxaspiro[3.5]nonan-7-one oxime 86b (1.45 g, 9.34 mmol) was dissolved in methanol (30 mL), and nickel chloride hexahydrate (2.22 g, 9.34 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 0.5 h and then cooled to -30 °C. Sodium borohydride (5.3 g, 140.1 mmol) was slowly added, and after addition, the reaction mixture was slowly warmed to room temperature and stirred overnight. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate, and the organic layer was concentrated to give the title compound 2-oxaspiro[3.5]nonan-7-amine 86c (pale yellow oil, 481 mg, 37.6% yield). LC-MS m/z(ESI) = 142.20 [M+1]
Step 3: Ethyl 4-((2-oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylate (86d)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (753 mg, 3.41 mmol) and potassium carbonate (940 mg, 6.81 mmol) were dissolved in acetonitrile (20 mL), and 8 oxabicyclo[3.2.1]octan-3-amine 86c (481 mg, 3.41 mmol) was added at 0 °C. The reaction mixture was warmed to room temperature and stirred for 20 h. The reaction was monitored by TLC. After completion of the reaction, water (30 mL) was added to the reaction mixture, and a solid was precipitated. The mixture was filtered, and the filter cake was washed 3 times with water and concentrated to obtain the title compound ethyl 4-((2-oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylate 86d (white solid, 640 mg, 57.6% yield). 'H NMR (400 MHz, DMSO) 6 8.61 (s, 1H), 8.29 (d, 1H), 4.34-4.29 (in, 4H), 4.24 (s, 2H), 3.98-3.89 (in, 1H), 2.05-1.92 (in, 2H), 1.86-1.75 (in, 2H), 1.65- 1.56 (in, 2H), 1.45 1.35 (in, 2H), 1.34-1.27 (in, 3H). LC-MS m/z(ESI) = 326.20 [M+1]
Step 4: 4-((2-Oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid (86e)
Ethyl 4-((2-oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylate 86d (640 mg, 1.96 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL of water, and lithium hydroxide (189 mg, 7.86 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 4, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 4-((2-oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5 carboxylic acid 86e (white solid, 518 mg, 88.5% yield). 'H NMR (400 MHz, DMSO) 6 13.74 (s, 1H), 8.57 (s, 1H), 8.48 (d, 1H), 4.32 (s, 2H), 4.24 (s, 2H), 3.97-3.84 (in, 1H), 1.99 (d, 2H), 1.86-1.76 (in, 2H), 1.66-1.54 (in, 2H), 1.42-1.26 (in, 2H). LC-MS m/z(ESI)= 298.10 [M+1]
Step 5: 2-Chloro-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one (86f)
4-((2-Oxaspiro[3.5]nonan-7-yl)amino)-2-chloropyrimidine-5-carboxylic acid 86e (518 mg, 1.74 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (175 mg, 1.74 mmol) and diphenylphosphoryl azide (479 mg, 1.74 mmol) were added. The reaction mixture was gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=5:1 to 1:10) to obtain the title compound 2-chloro 9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one 86f (white solid, 353 mg, 72.8% yield). H NMR (600 MHz, DMSO) 6 11.61 (s, 1H), 8.11 (s, 1H), 4.41 (s, 2H), 4.24 (s, 2H), 4.15-4.08 (m, 1H), 2.24-2.08 (m, 4H), 1.69 (d, 2H), 1.64-1.51 (m, 2H). LC-MS m/z(ESI)= 295.10 [M+1]
Step 6: 2-Chloro-7-methyl-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one (86g)
2-Chloro-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one 86f (353 mg, 1.18 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (150 mg, 1.18 mmol) and cesium carbonate (464 mg, 3.36 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, 30 mL of water was added, and the mixture was extracted with ethyl acetate. The organic phase was concentrated to afford the title compound 2 chloro-7-methyl-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one 86g (white solid, 313 mg, 85.5% yield). 'H NMR (400 MHz, DMSO) 6 8.34 (s, 1H), 4.41 (s, 2H), 4.25 (s, 2H), 4.22- 4.08 (m, 1H), 3.34 (s, 3H), 2.22-2.05 (m, 4H), 1.69 (d, 2H), 1.64-1.50 (m, 2H). LC-MS m/z(ESI) = 309.20 [M+1]
Step 7: 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(2-oxaspiro[3.5]nonan-7-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 86)
2-Chloro-7-methyl-9-(2-oxaspiro[3.5]nonan-7-yl)-7,9-dihydro-8H-purin-8-one 86g (60 mg, 0.194 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (33 mg, 0.194 mmol), cesium carbonate (127 mg, 0.39 mmol) and Brettphos Pd G3 (17 mg, 0.019 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 2-fluoro-5 methyl-4-((7-methyl-8-oxo-9-(2-oxaspiro[3.5]nonan-7-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide compound 86 (white solid, 21 mg, 24.7% yield). 'H NMR (400 MHz, DMSO-d) 68.50 (s, 1H), 8.17 (s, 1H), 7.74 (d, 1H), 7.55 (d, 1H), 7.47 (s, 1H), 7.37 (s, 1H), 4.32 (s, 2H), 4.23 (s, 2H), 4.13 (s, 1H), 3.31 (s, 3H), 2.27 (s, 3H), 2.16 (d, 4H), 1.72-1.64 (m, 2H), 1.53 (t, 2H). LC-MS m/z(ESI)= 441.20 [M+1]
Example 87 2-Fluoro-4-((9-(6-hydroxyspiro[3.3]heptan-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 87)
H2,N N'-' N= F N N H
OH Compound 87 O 0
HO Step 1 HO Step 2 I Step 3 N OH 'O N'Bc"O HHI CI N NH - CI N NH NBO NH2 HCI H
OH OH 87a 87b 87c 87d
H I0/ N N>= N~:N>=O Se Step 4 . CI=O CI Step 5 N N N CIH2N -N ApHNO F NN N H
OH OH OH
87e 87f Compound 87
Step 1: 6-Aminospiro[3.3]heptan-2-ol hydrochloride (87b)
tert-Butyl (6-hydroxyspiro[3.3]heptan-2-yl)carbamate 87a (2 g, 8.80 mmol) was dissolved in 4 M hydrogen chloride-1,4-dioxane (15 mL). The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate 1,4 dioxane solution to give the title compound 6-aminospiro[3.3]heptan-2-ol hydrochloride 87b (pale yellow solid, crude, 1.44 g, 100% yield). LC-MS m/z(ESI) = 163.10 [M+1]
Step 2: Ethyl 2-chloro-4-((6-hydroxyspiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylate (87c)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (2.92 g, 13.20 mmol) and 6 aminospiro[3.3]heptane-2-ol hydrochloride 87b (1.44 g, 8.80 mmol) were dissolved in acetonitrile (20 mL), and potassium carbonate (3.65 g, 26.40 mmol) was added with stirring. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. The reaction mixture was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate (v/v)=4:1) to obtain the title compound ethyl 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5 carboxylate 87c (white solid g, 1.36 g, 49.57% yield).
H NMR (400 MHz, DMSO-d6) 6 8.59 (s, 1H), 8.43 (d, 1H), 4.91 (s, 1H), 4.44-4.34 (m, 1H), 4.31 (q, 2H), 4.02-3.93 (m, 1H), 2.43-2.35 (m, 2H), 2.34-2.27 (m, 1H), 2.23 2.16 (m, 1H), 2.07-1.99 (m, 2H), 1.90-1.80 (m, 2H), 1.31 (t, 3H). LC-MS m/z(ESI) = 312.10 [M+1]
Step 3: 2-Chloro-4-((6-hydroxyspiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid (87d)
Ethyl 2-chloro-4-((6-hydroxyspiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylate 87c (1.0 g, 3.21 mmol) was dissolved in 10 mL of tetrahydrofuran and 10 mL of water, and lithium hydroxide (269 mg, 6.42 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 2 N hydrochloric acid, and a white solid was precipitated. The reaction mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to yield the title compound 2-chloro-4-((6 hydroxyspiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid 87d (760 mg, white solid, 83.52% yield). 'H NMR (400 MHz, DMSO-d6) 6 13.75 (s, 1H), 8.62 (d, 1H), 8.56 (s, 1H), 4.90 (s, 1H), 4.42-4.32 (m, 1H), 4.00-3.93 (m, 1H), 2.42-2.35 (m, 2H), 2.33-2.27 (m, 1H), 2.22 2.16 (m, 1H), 2.02-1.97 (m, 2H), 1.88-1.80 (m, 2H). LC-MS m/z(ESI)= 284.10 [M+1]
Step 4: 2-Chloro-9-(6-hydroxyspiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one (87e)
2-Chloro-4-((6-hydroxyspiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid 87d (760 mg, 2.68 mmol) was dissolved in dimethylacetamide (10 mL), and triethylamine (0.37 mL, 2.68 mmol) and diphenylphosphoryl azide (0.57 mL, 2.68 mmol) were added. The reaction mixture was gradually heated to 90 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, concentrated and dried to give the title compound 2-chloro-9-(6 hydroxyspiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one 87e (580 mg, white solid, 77.13% yield). H NMR (400 MHz, DMSO-d6) 6 11.60 (s, 1H), 8.10 (s, 1H), 4.96 (d, 1H), 4.70-4.60 (m, 1H), 2.92-2.87 (m, 2H), 2.46-2.43 (m, 2H), 2.32-2.21 (m, 3H), 1.92-1.87 (m, 2H). LC-MS m/z(ESI)= 281.10 [M+1]
Step 5: 2-Chloro-9-(6-hydroxyspiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one (87f)
2-Chloro-9-(6-hydroxyspiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one 87e (580 mg, 2.07 mmol) was dissolved in dimethylformamide (5 mL), and dimethyl sulfate (260 mg, 2.07 mmol) and cesium carbonate (1.3 g, 4.13 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to yield the title compound 2 chloro-9-(6-hydroxyspiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 87f (380 mg, white solid, 62.40% yield). 'H NMR (400 MHz, DMSO-d6) 68.33 (s, 1H), 4.97 (d, 1H), 4.70-4.66 (m, 1H), 4.05 3.96 (m, 1H), 3.33 (s, 3H), 2.95-2.87 (m, 2H), 2.51-2.43 (m, 1H), 2.31-2.25 (m, 3H), 1.93-1.87 (m, 2H). LC-MS m/z(ESI)= 295.10 [M+1]
Step 6: 2-Fluoro-4-((9-(6-hydroxyspiro[3.3]heptan-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 87)
2-Chloro-9-(6-hydroxyspiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 87f (130 mg, 0.44 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (222 mg, 1.32 mmol), cesium carbonate (311 mg, 0.88 mmol) and Brettphos Pd G3 (40 mg, 0.044 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 2-fluoro-4-((9 (6-hydroxyspiro[3.3]heptan-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) -methylbenzamide compound 87 (white solid, 35 mg, 18.62% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.51 (s, 1H), 8.18 (s, 1H), 7.85 (d, 1H), 7.55 (d, 1H), 7.47 (s, 1H), 7.38 (s, 1H), 4.97 (d, 1H), 4.72-4.63 (m, 1H), 4.03-3.94 (m, 1H), 3.29 (s, 3H), 2.99-2.90 (m, 2H), 2.46-2.40 (m, 1H), 2.29 (s, 3H), 2.27-2.13 (m, 3H), 1.92-1.83 (m, 2H). LC-MS m/z(ESI) = 427.20 [M+1]
Example 88 2-Fluoro-4-((9-(3-hydroxybicyclo[3.2.1]octane-8-yl)-7-methyl-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-5-methylbenzamide(compound88)
0 H2 N N N
F N "["N IN ) H
OH Compound 88
0 0 H
Boc'NH NH2 HCI N Step I N OH Step 4 NI O Step2 NNH1 NH C1 NN
O 0 0
88a 88b 88c 88d 88e
0 N N NN N N Step5 ,ICI NN Step 6 N Step 7 CI N N, Stp H2N01 - -N >szO F NN H 0
OH OH 88f 88g Compound 88
Step 1: 8-Aminobicyclo[3.2.1]octan-3-one hydrochloride (88b)
tert-Butyl (3-oxobicyclo[3.2.1]octan-8-yl)carbamate 88a (1.0 g, 4.18 mmol) was dissolved in 4 N dioxane hydrochloride (40 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was directly concentrated and dried to yield the target compound 8-aminobicyclo[3.2.1]octan-3-one hydrochloride 88b (white solid, 730 mg, 99.45% yield). 1H NMR (401 MHz, DMSO-d6) 6 8.77 (s, 2H), 3.36 (m, 2H), 2.80-2.77 (m, 2H), 2.16 2.13 (m, 2H), 1.87-1.85 (m, 2H), 1.43-1.42 (m, 2H). LC-MS m/z(ESI)= 176.20 [M+1]
Step 2: Ethyl 2-chloro-4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylate (88c)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (1.38 g, 6.26 mmol), 8 aminobicyclo[3.2.1]octan-3-one hydrochloride 88b (730 mg, 4.17 mmol) and potassium carbonate (1.73 g, 12.52 mmol) were dissolved in acetonitrile (20 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the solid was washed with a small amount of acetonitrile. The filtrates were combined and concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate (v/v)= 1:1) to obtain the target compound ethyl 2-chloro 4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylate 88c (white solid, 1.0 g, 74.01% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.86 (d, 2H), 8.68 (s, 1H), 4.36-4.30 (q, 2H), 4.20 4.16 (m, 1H), 2.66-2.65 (m, 2H), 2.56-2.55 (m, 2H), 2.21-2.17 (m, 2H), 1.97-1.93 (m, 2H), 1.53-1.33 (m, 2H), 1.31 (t, 3H). LC-MS m/z(ESI) = 324.10 [M+1]
Step 3: 2-Chloro-4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylic acid
(88d)
Ethyl 2-chloro-4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylate 88c (1.0 g, 3.09 mmol) was dissolved in 20 mL of tetrahydrofuran and 20 mL of water, and lithium hydroxide (259 mg, 6.18 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 2 N hydrochloric acid, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to give the title compound 2-chloro-4-((3 oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylic acid 88d (white solid, 800 mg, 87.59% yield). H NMR (400 MHz, DMSO-d6) 6 13.93 (s, 1H), 9.20 (d, 2H), 8.64 (s, 1H), 4.20-4.16 (m, 1H), 2.66-2.64 (m, 2H), 2.54-2.53 (m, 2H), 2.21-2.16 (m, 2H), 1.96-1.93 (m, 2H), 1.53-1.47 (m, 2H). LC-MS m/z(ESI)= 296.10 [M+1]
Step 4: 2-Chloro-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one (88e)
2-Chloro-4-((3-oxobicyclo[3.2.1]octan-8-yl)amino)pyrimidine-5-carboxylic acid 88d (800 mg, 2.71 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (0.37 mL, 2.7 mmol) and diphenylphosphoryl azide (0.58 mL, 2.7 mmol) were added. The reaction mixture was gradually heated to 90 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, and dried to obtain the title compound 2-chloro-9-(3-oxobicyclo[3.2.1]octan-8 yl)-7,9-dihydro-8H-purin-8-one 88e (white solid, 630 mg, 94.71% yield). H NMR (400 MHz, DMSO-d6) 6 11.66 (s, 1H), 8.13 (s, 1H), 4.01-3.99 (m, 1H), 3.65 3.64 (m, 2H), 2.66-2.61 (m, 2H), 2.12-2.08 (m, 2H), 1.89-1.86 (m, 2H), 1.54-1.49 (m, 2H). LC-MS m/z(ESI) = 293.00 [M+1]
Step 5: 2-Chloro-7-methyl-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one (88f)
2-Chloro-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one 88e (630 mg, 2.15 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (0.2 mL, 2.15 mmol) and cesium carbonate (1.4 g, 4.3 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to yield the title compound 2 chloro-7-methyl-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one 88f (white solid, 490 mg, 74.22% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.37 (s, 1H), 4.03-4.01 (m, 1H), 3.66-3.65 (m, 2H), 3.36 (s, 3H), 2.65-2.59 (m, 2H), 2.13-2.08 (m, 2H), 1.89-1.87 (m, 2H), 1.55-1.50 (m, 2H). LC-MS m/z(ESI) = 307.10 [M+1]
Step 6: 2-Chloro-9-(3-hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-7,9-dihydro-8H-purin-8 one (88g)
2-Chloro-7-methyl-9-(3-oxobicyclo[3.2.1]octan-8-yl)-7,9-dihydro-8H-purin-8-one 88f (490 mg, 1.6 mmol) was dissolved in tetrahydrofuran (30 mL) and methanol (30 mL), and sodium borohydride (181 mg, 4.79 mmol) was added at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to yield the title compound 2-chloro-9-(3 hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 88g (white solid, 380 mg, 77.04% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.37 (s, 1H), 4.28-4.20 (m, 1H), 3.74-3.66 (m, 2H), 3.45-3.43 (m, 2H), 3.37 (s, 3H), 2.14-2.07 (m, 1H), 1.74-1.50 (m, 5H), 1.28-1.22 (m, 2H). LC-MS m/z(ESI) = 309.10 [M+1]
Step 7: 2-Fluoro-4-((9-(3-hydroxybicyclo[3.2.1]octane-8-yl)-7-methyl-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-5-methylbenzamide (compound 88)
2-Chloro-9-(3-hydroxybicyclo[3.2.1]octan-8-yl)-7-methyl-7,9-dihydro-8H-purin-8 one 88g (100 mg, 0.32 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (164 mg, 0.96 mmol), cesium carbonate (228 mg, 0.64 mmol) and Brettphos Pd G3 (29 mg, 0.032 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 2-fluoro-4-((9 (3-hydroxybicyclo[3.2.1]octane-8-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide compound 88 (white solid, 83 mg, 59.20% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.49 (d, 1H), 8.21 (s, 1H), 7.75 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.40 (s, 1H), 4.31-4.26 (m, 1H), 3.75-3.71 (m, 1H), 3.62 (t, 1H), 3.52 (s, 2H), 3.34 (s, 3H), 2.28 (s, 3H), 1.72-1.62 (m, 4H), 1.59-1.50 (m, 2H), 1.35 (t, 2H). LC-MS m/z(ESI) = 441.20 [M+1]
Example 89 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(2-oxaspiro[3.3]heptan-6-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 89)
0 H2 N N N F N<N N H
Compound 89
NNN Step 2 hi OH HH Step 3 N tp1 ISteptp CI CIIN NH ~ INN CI N- CI
IA 89a 89b
H 0
CI N N C I n11Nz H
o o 0 89c 89d Compound 89
Step 1: Ethyl4-((2-oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylate(89a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (4.43 g, 20.05 mmol), 2 oxaspiro[3.3]heptan-6-amine hydrochloride (2.0 g, 13.37 mmol) and potassium carbonate (5.54 g, 40.10 mmol) were dissolved in acetonitrile (60 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the solid was washed with a small amount of acetonitrile. The filtrates were combined and concentrated, and the crude product was purified by column chromatography (petroleum ether:ethyl acetate (v/v) = 1:1) to obtain the target compound ethyl 4-((2 oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylate 89a (white solid, 3.0 g, 75.38% yield). 1H NMR (400 MHz, DMSO-d6) 68.60 (s, 1H), 8.47 (d, 1H), 4.63 (s, 2H), 4.49 (s, 2H), 4.39-4.33 (m, 1H), 4.30-4.27 (q, 2H), 2.66-2.61 (m, 2H), 2.31-2.26 (m, 2H), 1.30 (t, 3H). LC-MS m/z(ESI) = 298.10 [M+1]
Step 2: 4-((2-Oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylic acid (89b)
Ethyl 4-((2-oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylate 89a (3.0 g, 10.08 mmol) was dissolved in tetrahydrofuran/water (30 mL/30 mL), and lithium hydroxide (845 mg, 20.15 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 2 N hydrochloric acid, and a white solid was precipitated. The reaction mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to obtain the title compound 4-((2-oxaspiro[3.3]heptan-6 yl)amino)-2-chloropyrimidine-5-carboxylic acid 89b (white solid, 1.8 g, 66.24% yield). 'H NMR (400 MHz, DMSO-d6) 613.76 (s, 1H), 8.64 (d, 1H), 8.57 (s, 1H), 4.64 (s, 2H), 4.49 (s, 2H), 4.40-4.30 (m, 1H), 2.67-2.61 (m, 2H), 2.28-2.23 (m, 2H). LC-MS m/z(ESI) = 270.20 [M+1]
Step 3: 2-Chloro-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one (89c)
4-((2-Oxaspiro[3.3]heptan-6-yl)amino)-2-chloropyrimidine-5-carboxylic acid 89b (1.8 g, 6.67 mmol) was dissolved in dimethylacetamide (40 mL), and triethylamine (0.92 mL, 6.67 mmol) and diphenylphosphoryl azide (1.4 mL, 6.67 mmol) were added. The reaction mixture was gradually heated to 90 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, and dried to give 2-chloro-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one 89c (white solid, 1.3 g, 73.03% yield). 'H NMR (400 MHz, DMSO-d6) 6 11.61 (s, 1H), 8.11 (s, 1H), 4.71-4.63 (m, 5H), 3.02 2.94 (m, 2H), 2.69-2.66 (m, 2H). LC-MS m/z(ESI) = 267.10 [M+1]
Step 4: 2-Chloro-7-methyl-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one (89d)
2-Chloro-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one 89c (1.3 g, 4.87 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (0.46 mL, 4.87 mmol) and cesium carbonate (3.18 g, 9.75 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to obtain the title compound 2-chloro 7-methyl-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one 89d (white solid, 875 mg, 63.94% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.35 (s, 1H), 4.76-4.69 (m, 1H), 4.66-4.63 (m, 4H), 3.33 (s, 3H), 3.01-2.95 (m, 2H), 2.71-2.66 (m, 2H). LC-MS m/z(ESI)= 281.10 [M+1]
Step 5: 2-Fluoro-5-methyl-4-((7-methyl-8-oxo-9-(2-oxaspiro[3.3]heptan-6-yl)-8,9-dihydro 7H-purin-2-yl)amino)benzamide (compound 89)
2-Chloro-7-methyl-9-(2-oxaspiro[3.3]heptan-6-yl)-7,9-dihydro-8H-purin-8-one 89d (200 mg, 0.71 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (359 mg, 2.13 mmol), cesium carbonate (502 mg, 1.42 mmol) and Brettphos Pd G3 (65 mg, 0.071 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 2-fluoro-5 methyl-4-((7-methyl-8-oxo-9-(2-oxaspiro[3.3]heptan-6-yl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide compound 89 (white solid, 58 mg, 19.82% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.49 (s, 1H), 8.19 (s, 1H), 7.80 (d, 1H), 7.56 (d, 1H), 7.47 (s, 1H), 7.41 (s, 1H), 4.67 (s, 3H), 4.52 (s, 2H), 3.30 (s, 3H), 3.08-3.02 (m, 2H), 2.65-2.57 (m, 2H), 2.27 (s, 3H). LC-MS m/z(ESI) = 413.20 [M+1]
Example 90 4-((9-(4-Cyanobicyclo[2.2.2]octan-1-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide(compound90)
0 H2 N N NNO F NNN N H
Compound 90 CN H - Step 1 0 0 Step 2 0 Step 3
HO 2H HN - O N- Q O
90a 90b 90c
0 0
N O/~ N N OH O Step 4 Step 5 / Step 6 CINNH N NH CI N NH C NC OH NC NH 2HCI
90d 90e g0f CN 90g CN
H 0
Step 7 1C OStep 8 NN CI NN/ O Step 9 H 2NN O CI N ~F H NN >sz
90h CN 90i CN Compound 90 CN
Step 1: Methyl 4-carbamoylbicyclo[2.2.2]octane-1-carboxylate (90b)
4-(Methoxycarbonyl)bicyclo[2.2.2]octane-1-carboxylic acid 90a (10.0 g, 47.12 mmol) was dissolved in anhydrous dichloromethane (200 mL), and 2-(7-azabenzotriazole) N,N,N',N'-tetramethylurea hexafluorophosphate (18.81 g, 49.47 mmol) was added in an ice bath. The mixture was stirred for 30 min with the temperature maintained. N,N Diisopropylethylamine (24.62 mL, 141.35 mmol) was added, and then solid ammonium chloride (3.78 g, 70.67 mmol) was slowly added several times. The reaction mixture was gradually warmed to room temperature and stirred overnight. The reaction was monitored by TLC. After completion of the reaction, 100 mL of 0.5 N hydrochloric acid solution was added, the organic phase was separated, successively with water and saturated brine (100 mL each), dried and concentrated to give the title compound methyl 4-carbamoylbicyclo[2.2.2]octane-1-carboxylate 90b (white solid, 21 g, crude). 'H NMR (400 MHz, DMSO-d6) 6 6.94 (s, 1H), 6.73 (s, 1H), 3.57 (s, 3H), 1.72-1.63 (m, 12 H). LC-MS m/z(ESI)= 212.10 [M+1]
Step 2: Methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate (90c)
Methyl 4-carbamoylbicyclo[2.2.2]octane-1-carboxylate 90b (21 g, 99.40 mmol) was dissolved in dichloromethane (200 mL), and pyridine (16.02 mL, 198.81 mmol) and trifluoroacetic anhydride (21.00 mL, 149.10 mmol) were added in the ice bath. The reaction mixture was stirred for 1 h with the temperature maintained. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered directly, and the filter cake was washed with 100 mL of dichloromethane. The organic phases were combined, then extracted with 1 N hydrochloric acid solution, water and saturated brine (100 mL each), dried, and concentrated, and the residue was purified by column chromatography to obtain the target product methyl 4 cyanobicyclo[2.2.2]octane-1-carboxylate 90c (white solid, 5.3 g, 58.21% yield over two steps). H NMR (400 MHz, CDCl3-d6) 6 3.66 (s, 3H), 1.98-1.94 (in, 6H), 1.86-1.82 (in, 6H). LC-MS m/z(ESI)= 194.10 [M+1]
Step 3: 4-Cyanobicyclo[2.2.2]octane-1-carboxylic acid (90d)
Methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate 90c (5.3 g, 27.43 mmol) was dissolved in 50 mL of tetrahydrofuran and 50 mL of water, and lithium hydroxide (1.73 g, 41.14 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 2 N hydrochloric acid, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to obtain the title compound 4-cyanobicyclo[2.2.2]octane-1-carboxylic acid 90d (white solid, 5.0 g, crude). 'H NMR (400 MHz, DMSO-d6) 6 12.23 (s, 1H), 1.89-1.85 (in, 6H), 1.72 -1.68 (in, 6H). LC-MS m/z(ESI)= 180.10 [M+1]
Step 4: 4-Aminobicyclo[2.2.2]octane-1-carbonitrile hydrochloride (90e)
4-Cyanobicyclo[2.2.2]octane-1-carboxylic acid 90d (5.0 g, 27.90 mmol) was dissolved in toluene (60 mL), and diphenylphosphoryl azide (6.01 mL, 27.90 mmol) and triethylamine (3.88 mL, 27.90 mmol) were added in an ice bath. The reaction mixture was stirred at room temperature for 1 h, then heated to 90 °C and stirred for another 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature and slowly poured into 100 mL of 1 N hydrochloric acid solution, and a large amounts of solid was precipitated. The solid was collected by filtration, slurried with ethyl acetate (150 mL), and dried in vacuo to yield the title compound 4-aminobicyclo[2.2.2]octane-1-carbonitrile hydrochloride 90e (white solid, 7.3 g, crude). 'H NMR (401 MHz, DMSO-d6) 6 8.45 (s, 2H), 2.01-1.97 (in, 6H), 1.81-1.76 (in, 6H). LC-MS m/z(ESI) = 187.10 [M+1]
Step 5: Ethyl 2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl) amino)pyrimidine-5-carboxylate (90f)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (8.74 g, 39.53 mmol), 4 aminobicyclo[2.2.2]octane-1-carbonitrile hydrochloride 90e (7.38 g, 39.53 mmol) and potassium carbonate (21.85 g, 158.13 mmol) were dissolved in acetonitrile (200 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the solid was washed with a small amount of acetonitrile. The filtrates were combined and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1:1) to obtain the title compound ethyl 2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl) amino)pyrimidine-5-carboxylate f (white solid, 4.0 g, 30.23% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.63 (s, 1H), 8.29 (s, 1H), 4.30 (q, 2H), 2.10-2.00 (m, 12H), 1.30 (t, 3H). LC-MS m/z(ESI)= 335.10 [M+1]
Step 6: 2-Chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid (90g)
Ethyl 2-chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl) amino)pyrimidine-5-carboxylate f (4 g, 11.95 mmol) was dissolved in 50 mL of tetrahydrofuran and 50 mL of water, and lithium hydroxide (1.01 g, 23.90 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 2 N hydrochloric acid, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to afford the title compound 2-chloro-4-((4 cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid 90g (white solid, 3.6 g, 98.23% yield). H NMR (600 MHz, DMSO-d6) 6 13.85 (s, 1H), 8.59 (s, 1H), 8.56 (s, 1H), 2.07-1.98 (m, 12H). LC-MS m/z (ESI) = 307.10 [M+1]
Step 7: 4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile (90h)
2-Chloro-4-((4-cyanobicyclo[2.2.2]octan-1-yl)amino)pyrimidine-5-carboxylic acid g (3.8 g, 12.39 mmol) was dissolved in dimethylacetamide (50 mL), and triethylamine (1.72 mL, 12.39 mmol) and diphenylphosphoryl azide (2.67 mL, 12.39 mmol) were added. The reaction mixture was gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, and dried to give the title compound 4-(2-chloro-8-oxo-7,8 dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile 90h (white solid, 3.3 g, 87.7% yield). H NMR (400 MHz, DMSO-d6) 6 11.61 (s, 1H), 8.09 (s, 1H), 2.48-2.40 (m, 6H), 2.09 2.03 (m, 6H). LC-MS m/z(ESI) = 304.20 [M+1]
Step 8:
4-(2-Chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1 carbonitrile (90i)
4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carbonitrile 90h (3.3 g, 11.43 mmol) was dissolved in dimethylformamide (50 mL), and dimethyl sulfate (1.44 g, 11.43 mmol) and cesium carbonate (5.59 g, 17.15 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to obtain the title compound 4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1 carbonitrile 90i (white solid, 3.1 g, 89.59% yield). 1H NMR (401 MHz, DMSO-d6) 6 8.33 (s, 1H), 3.29 (s, 3H), 2.48-2.41 (m, 6H), 2.10
2.04 (m, 6H). LC-MS m/z(ESI)= 318.20 [M+1]
Step 9: 4-((9-(4-Cyanobicyclo[2.2.2]octan-1-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 90)
4-(2-Chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1 carbonitrile 90i (200 mg, 0.63 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (318 mg, 1.89 mmol), cesium carbonate (444 mg, 1.26 mmol) and Brettphos Pd G3 (57 mg, 0.063 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 30/1) to obtain the title compound 4-((9-(4-cyanobicyclo[2.2.2]octan-1-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide compound 90 (white solid, 18 mg, 6.4% yield). 1H NMR (401 MHz, DMSO-d6) 6 10.08 (d, 1H), 8.22-8.17 (s, 1H), 7.88-7.53 (d, 1H), 7.48-7.29 (d, 1H), 6.31-6.29 (s, 1H), 5.82 (s, 1H), 3.26 (d, 3H), 2.34-2.26 (m, 6H), 2.06 (d, 5H), 1.96 (dd, 4H). LC-MS m/z(ESI) = 450.20 [M+1]
Example 91 2-Fluoro-4-((9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide(compound91)
0/ H2 N N N
F N N N H O-0H
Compound 91
0 OH OH N1 O Step I Step 2 Step 3 Step 4 CI N NH N'Boc NBoc N2C H H NH 2HCI OH
91a 91b 91c 91d
0 H N0/
StepN5 C NOH N S CO Step H 2N N N
H H OH OH OH
91e 91f 91g Compound 91
Step 1: tert-Butyl (3-hydroxy-3-methylcyclohexyl)carbamate (91b)
tert-Butyl (3-oxocyclohexyl)carbamate 91a (5.00 g, 23.4 mmol) was dissolved in tetrahydrofuran (80mL). The solution was pre-cooled at-78 °C, and then 1.6 M methyllithium-diethyl ethersolution (61.5 mL, 98.28 mmol) was slowly added dropwise over 45min.The reaction mixture was stirred at -78°C for 1h, and then the remaining 1.6 Mmethyllithium-diethyl ether solution (61.5 mL, 98.28 mmol) was added. The reaction mixture was stirred at -78°C for another hour. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride solution, and water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 8:1) to obtainthe titlecompound tert-butyl (3-hydroxy-3-methylcyclohexyl)carbamate 91b (white solid, 2.07 g, 38.53% yield). 1HNMR (400 MHz, Chloroform-d)64.30 (s, 1H), 3.68 (s, 1H), 1.90 (d, 3H), 1.67 (q, 1H), 1.59-1.48 (i,2H), 1.37 (s, 9H), 1.19-1.12 (in,4H), 1.06 (t, 1H), 0.90 (qd, 1H). LC-MS m/z(ESI)= 230.20 [M+1]
Step 2: 3-Amino-i-methylcyclohexan-1-olhydrochloride (91c)
tert-Butyl (3-hydroxy-3-methylcyclohexyl)carbamate 91b (2.07g, 9.03 mmol) was dissolved in4 M hydrogenachloride-1,4-dioxane (10 mL). The reaction mixture was stirred at roomtemperature. Thereaction was monitored byhTLC. Aftercompletion of the reaction, the reaction mixture was concentratedtoevaporate1,4-dioxanesolution to give the title compound3-amino-1-methylcyclohexan-1-olhydrochloride91c(pale yellow solid, crude, 1.49g). LC-MS m/z(ESI) = 166.10 [M+1]
Step 3: Ethyl 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylate (91d)
Ethyl2,4-dichloropyrimidine-5-carboxylateA (2.99 g,13.54 mmol) and 3-amino-1 methylcyclohexan-1-olhydrochloride 91c(1.49 g, 9.03 mmol) weredissolved in acetonitrile(20mL),andpotassiumcarbonate (3.74 g, 27.08 mmol) was addedwith stirring. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate (v/v) = 4:1) to obtain the title compound ethyl 2-chloro-4-((3 hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylate 91d (white solid, 2.23 g, 78.74% yield). H NMR (400 MHz, Chloroform-d) 6 8.57 (s, 1H), 8.19 (d, 1H), 4.43-4.32 (m, 1H), 4.27 (q, 2H), 2.07-1.95 (m, 2H), 1.83-1.69 (m, 2H), 1.64-1.56 (m, 2H), 1.31 (t, 4H), 1.26 (d, 1H), 1.21 (s, 3H), 1.10-1.05 (m, 1H). LC-MS m/z(ESI) = 314.10 [M+1]
Step 4: 2-Chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (91e)
Ethyl 2-chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylate 91d (2.23 g, 7.11 mmol) was dissolved in tetrahydrofuran/water (15 mL/15 mL), and lithium hydroxide (895 mg, 21.32 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 3-4 with 2 N hydrochloric acid, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to give the title compound 2-chloro-4-((3-hydroxy-3 methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 91e (white solid, 1.6 g, 78.79% yield). LC-MS m/z(ESI) = 286.10 [M+1]
Step 5: 2-Chloro-9-(3-hydroxy-3-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (91f)
2-Chloro-4-((3-hydroxy-3-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 91e (1.38 g, 4.83 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (0.67 mL, 4.83 mmol) and diphenylphosphoryl azide (1.04 mL, 4.83 mmol) were added. The reaction mixture was gradually heated to 90 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water. The solid was collected by filtration and washed 3 times with water, and concentrated and dried to obtain the target compound 2-chloro-9-(3 hydroxy-3-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 91f (white solid, 1.35 g, 98.86% yield). H NMR (400 MHz, DMSO-d6) 6 11.60 (s, 1H), 8.10 (s, 1H), 4.62-4.54 (m, 1H), 4.35 (s, 1H), 2.19 (t, 1H), 2.10-1.99 (m, 1H), 1.70-1.53 (m, 5H), 1.30-1.22 (m, 1H), 1.16 (s, 3H). LC-MS m/z(ESI)= 283.10 [M+1]
Step 6: 2-Chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (91g)
2-Chloro-9-(3-hydroxy-3-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one 91f (1.35 g,
4.77 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (0.45 mL, 4.77 mmol) and cesium carbonate (1.56 g, 4.77 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and a solid was precipitated. The solid was collected by filtration to give the title compound 2 chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 91g (white solid, 873 mg, 61.61% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.34 (s, 1H), 4.66-4.58 (m, 1H), 4.37 (s, 1H), 3.35 (s, 3H), 2.20 (t, 1H), 2.11-2.00 (m, 1H), 1.72-1.54 (m, 5H), 1.32-1.24 (m, 1H), 1.17 (s, 3H). LC-MS m/z(ESI) = 297.10 [M+1]
Step 7: 2-Fluoro-4-((9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 91)
2-Chloro-9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 91g (200 mg, 0.67 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (338 mg, 2.01 mmol), cesium carbonate (473 mg, 1.34 mmol) and Brettphos Pd G3 (61 mg, 0.067 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 2-fluoro-4-((9 (3-hydroxy-3-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) -methylbenzamide compound 91 (white solid, 140 mg, 48.80% yield). 'H NMR (400 MHz, DMSO-d) 68.50 (s, 1H), 8.18 (s, 1H), 7.96 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.39 (d, 1H), 4.67-4.58 (m, 1H), 4.33 (s, 1H), 3.32 (s, 3H), 2.35 (t, 1H), 2.30 (s, 3H), 2.15-2.01 (m, 1H), 1.80-1.63 (m, 3H), 1.58 (d, 2H), 1.41-1.22 (m, 1H), 1.18 (s, 3H). LC-MS m/z(ESI) = 429.20 [M+1]
Example 92 4-((9-(4-Cyano-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide(compound92)
0
H2 N N N
H
Compound 92 NC
N ~ of"aCN Step1 -tp Step CN Step 2 CN Step 3 CIN NHO Step4 0 0( C H 2N'C:
NC 92a 92b 92c 92d
0 H /0
C OH Step 5 CSt Step6 , H 2N N NH . IN IN.. CIN F N N N)= H
NC NC NC92f 92g Compound 92C 92e
Step 1: 1-Methyl-4-oxocyclohexane-1-carbonitrile (92b)
1,4-Dioxaspiro[4.5]decane-8-carbonitrile 92a (5.00 g, 29.9 mmol) was dissolved in THF (60 mL), and lithium bistrifluoromethanesulfonimide (9.87 mL, 34.38 mmol) was slowly added dropwise at 0 °C. The reaction mixture was stirred for 1 h, and then iodomethane (4.24 g, 29.9 mmol) was slowly added. The reaction mixture was stirred for another hour. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was extracted twice with saturated ammonium chloride solution, bromine water and ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to get an intermediate 8-methyl-1,4 dioxaspiro[4.5]decane-8-carbonitrile, which was then dissolved in THF (60 mL). 3 M HCl (60 mL) was added to the reaction mixture, which was then heated to 50 °C and stirred for 5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, the pH was adjusted to 7-8 with 3 M NaOH, and extracted 3 times with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the title compound 1-methyl 4-oxocyclohexane-1-carbonitrile 92b (yellow liquid, 3.48 g, 84.84% yield). LC-MS m/z(ESI) = 138.10 [M+i]
Step 2: 4-Amino-i-methylcyclohexane-1-carbonitrile (92c)
I-Methyl-4-oxocyclohexane-1-carbonitrile 92b (3.48 g, 25.37 mmol) was dissolved in absolute methanol (50 mL), and ammonium formate (16.00 g, 253.70 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 4 h, and then sodium cyanoborohydride (1.88 g, 30.44 mmol) was added. The reaction mixture was stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, the pH of the reaction mixture was adjusted to 1-2 with 2 N HCl, stirred for 0.5 h, then extracted twice with ethyl acetate, the pH was adjusted to about 10 with 2 N NaOH solution, and extracted 3 times with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the title compound 4-amino-i methylcyclohexane--carbonitrile 92c (pale yellow liquid, 2.80 g, 79.85% yield). LC-MS m/z(ESI)= 139.10 [M+i]
Step 3: Ethyl 2-chloro-4-((4-cyano-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate (92d)
Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1A (6.72 g, 30.39 mmol) was dissolved in acetonitrile (30 mL), and potassium carbonate (8.40 g, 60.78 mmol) was added with stirring at 0 °C, followed by the slow dropwise addition of a solution of 4-amino-1 methylcyclohexane-1-carbonitrile 92c (2.80 g, 20.26 mmol) in acetonitrile (10 mL). The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 20:1) to obtain the title compound ethyl 2-chloro-4-((4-cyano-4 methylcyclohexyl)amino)pyrimidine-5-carboxylate 92d (white solid, 2.63 g, 40.22% yield). H NMR (400 MHz, Chloroform-d) 6 8.68 (d, 1H), 4.38 (p, 2H), 2.23-1.82 (m, 4H), 1.79-1.49 (m, 6H), 1.46-1.36 (m, 6H). LC-MS m/z(ESI) = 323.10 [M+1]
Step 4: 2-Chloro-4-((4-cyano-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (92e)
Ethyl 2-chloro-4-((4-cyano-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate 92d (2.63 g, 8.15 mmol) was dissolved in tetrahydrofuran/water (20 mL/10 mL), and lithium hydroxide monohydrate (1.00 g, 24.45 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to about 3-4 with 2 N HCl, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to obtain the title compound 2 chloro-4-((4-cyano-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 92e (white solid, 2.20 g, 91.59% yield). 'H NMR (400 MHz, DMSO-d6) 6 13.75 (s, 1H), 8.59 (s, 1H), 8.52 (d, 1H), 3.97 (m, 1H), 2.06-1.92 (m, 4H), 1.63-1.47 (m, 4H), 1.35 (s, 3H). LC-MS m/z(ESI)= 295.10 [M+1]
Step 5: 4-(2-Chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)-1-methylcyclohexane-1-carbonitrile (92f)
2-Chloro-4-((4-cyano-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid 92e (2.20 g, 7.46 mmol) was dissolved in N,N-dimethylacetamide (15 mL), and triethylamine (1.04 mL, 7.46 mmol) and diphenylphosphoryl azide (1.61 mL, 7.46 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted three times with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 80/1) to obtain the title compound 4-(2-chloro-8-oxo-8,9-dihydro-7h-purin-9-yl)-1 methylcyclohexane-1-carbonitrile 92f (white solid, 0.9 g, 41.35% yield).
H NMR (400 MHz, DMSO-d6) 6 11.64 (s, 1H), 8.13 (s, 1H), 4.31-4.20 (m, 1H), 2.45 2.33 (m, 2H), 2.01-1.94 (m, 4H), 1.71-1.64 (m, 2H), 1.49 (s, 3H). LC-MS m/z(ESI)= 292.10 [M+1]
Step 6: 4-(2-Chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)-1-methylcyclohexane-1 carbonitrile (92g)
4-(2-chloro-8-oxo-8,9-dihydro-7h-purin-9-yl)-1-methylcyclohexane-1-carbonitrile 92f (0.90 g, 3.08 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dimethyl sulfate (0.29 mL, 3.08 mmol) and cesium carbonate (1.51 g, 4.62 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred, and a white solid was precipitated. The solid was washed 3 times with water and petroleum ether and collected by filtration to give the title compound 4-(2-chloro-7-methyl-8-oxo-8,9 dihydro-7h-purin-9-yl)-1-methylcyclohexane-1-carbonitrile 92g (white solid, 0.85 g, 90.26% yield). H NMR (400 MHz, DMSO-d6) 6 8.35 (s, 1H), 4.35-4.20 (m, 1H), 3.36 (s, 3H), 2.48 2.31 (m, 2H), 2.06-1.95 (m, 3H), 1.83-1.76 (m, 1H), 1.71-1.57 (m, 2H), 1.48 (s, 1H), 1.36 (s, 2H). LC-MS m/z(ESI) = 306.10 [M+1]
Step 7: 4-((9-(4-Cyano-4-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 92)
2-Chloro-(9-(3-hydroxy-3-methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one 92g (200 mg, 0.67 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (338 mg, 2.01 mmol), cesium carbonate (473 mg, 1.34 mmol) and Brettphos Pd G3 (61 mg, 0.067 mmol) were dissolved in dioxane, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was s monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 2-fluoro-4-((9 (3-hydroxyl-3-methylcyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino) -methylbenzamide compound 92, which was resolved by SFC to give compound 92 1 (white solid, 12 mg, 4.1% yield, RT = 3.58 min) and compound 92-2 (white solid, 22 mg, 7.5% yield, RT = 4.17 min). (OZ), mobile phase: C02/ethanol = 70/30; column temperature: 35 °C; column pressure: 80 bar; flow rate: 1 mL/min; detector signal channel: 215 nm@4.8 nm; diode array detector at a wavelength of 200-400 nm. Compound 92-1: H NMR (400 MHz, DMSO-d) 68.60 (s, 1H), 8.18 (s, 1H), 7.65 (d, 1H), 7.57 (d, 1H), 7.47 (s, 1H), 7.38 (s, 1H), 4.27-4.16 (m, 1H), 2.45-2.31 (m, 5H), 2.26 (s, 3H), 1.97-1.89 (m, 4H), 1.69-1.59 (m, 2H), 1.32 (s, 3H). LC-MS m/z(ESI) = 438.20 [M+1]
Compound 92-2: H NMR (400 MHz, DMSO-d) 68.32 (s, 1H), 8.19 (s, 1H), 7.91 (d, 1H), 7.53 (d, 1H), 7.44 (s, 1H), 7.27 (s, 1H), 4.28-4.18 (m, 1H), 2.26 (s, 3H), 2.08-2.00 (m, 2H), 1.85-1.77 (m, 2H), 1.60-1.49 (m, 3H), 1.37 (s, 3H), 1.26-1.13 (m, 4H). LC-MS m/z(ESI) = 438.20 [M+1]
Example 93 2-Fluoro-4-((9-(trans-3-hydroxycyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 93)
0
H 2N N N F: N -NN N>= H
Compound 93 OH
0 0 0 Stpi N N 0'O6 N N OH N Step3 Step t CIN Step2 : .. Hj NHNH Stp CI N CI
OTBDPS OTBDPS OH IA 93a 93b 93c
H 0 0 N Step 6 H 2 N N N Step 7 H 2N N N NO N N H H
93d OTBDPS 93e OTBDPS 93f OTBDPS Compound 93 OH
Step 1: Ethyl 2-chloro-4-((trans-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate (93a)
trans-3-Aminocyclobutane-1-ol hydrochloride (3.00 g, 34.48 mmol) was dissolved in acetonitrile (25 mL), and potassium carbonate (14.30 g, 103.45 mmol) and ethyl 2,4 dichloro-5-pyrimidinecarboxylate 1A (11.43 g, 51.72 mmol) were added with stirring at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=2:1) to obtain the title compound ethyl 2-chloro-4-(((1r,3r)-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate 93a (white solid, 2.9 g, 30.95% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.61 (s, 1H), 8.48 (d, 1H), 5.17-5.11 (m, 1H), 4.61 4.51 (m, 1H), 4.36-4.27 (m, 3H), 2.32-2.20 (m, 4H), 1.32 (t, 3H). LC-MS m/z(ESI) = 272.00 [M+1]
Step 2: Ethyl 4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyri midine-5-carboxylate (93b)
Ethyl 2-chloro-4-((trans-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate 93a (3.10 g, 11.41 mmol) was dissolved in dichloromethane (20 mL), and tert butyldiphenylchlorosilane (4.45 mL, 17.11 mmol) and imidazole (1.94 g, 28.52 mmol) were added, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 10:1) to obtain the title compound ethyl 4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2 chloropyri-midine-5-carboxylate 93b (colorless liquid, 4.18 g, 64.26% yield). LC-MS m/z(ESI)= 510.20 [M+1]
Step 3: 4-((trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5 carboxylic acid (93c)
Ethyl 4-((trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyri midine-5-carboxylate 93b (4.18 g, 8.19 mmol) was dissolved in tetrahydrofuran/water (30 mL/15 mL), and lithium hydroxide monohydrate (1.03 g, 24.58 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to 3-4 with 2 N HCl, and water was added and the organics were extracted two times with ethyl acetate. The organic layer was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20:1) to obtain the title compound 4-((trans-3 ((tert-butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5-carboxylic acid 93c (white solid, 3.87 g, 97.97% yield). LC-MS m/z(ESI) = 482.20 [M+1]
Step 4: 9-(trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9-dihydro-8H-purin 8-one (93d)
4-((trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)amino)-2-chloropyrimidine-5 carboxylic acid 93c (3.87 g, 8.03 mmol) was dissolved in N,N-dimethylacetamide (38 mL), and triethylamine (1.11 mL, 8.03 mmol) and diphenylphosphoryl azide (1.73 mL, 8.03 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1) to obtain the title compound 9-((1r,3r)-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9 dihydro-8H-purin-8-one 93d (white solid, 1.81 g, 47.06% yield). 'H NMR (400 MHz, DMSO-d6) 6 11.59 (s, 1H), 8.09 (s, 1H), 7.65-7.62 (m, 2H), 7.62 7.60 (m, 2H), 7.47-7.39 (m, 6H), 5.05-4.97 (m, 1H), 4.96-4.89 (m, 1H), 2.98-2.84 (m, 2H), 2.54-2.46 (m, 2H), 1.02 (s, 9H). LC-MS m/z(ESI) = 479.20 [M+1]
Step 5: 9-(trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl-7,9-dihydro 8H-purin-8-one (93e)
9-(trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7,9-dihydro-8H-purin
8-one 93d (1.81 g, 3.78 mmol) was dissolved in N,N-dimethylformamide (20 mL), and dimethyl sulfate (0.36 mL, 3.78 mmol) and cesium carbonate (2.47 g, 7.57 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred and extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 3:1) to obtain the title compound 9-(trans-3-((tert-butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl 7,9-dihydro-8H-purin-8-one 93e (white solid, 1.80 g, 96.62% yield). LC-MS m/z(ESI)= 493.20 [M+1]
Step 6: 4-((9-(trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)-7-methyl-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide (93f)
9-(trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)-2-chloro-7-methyl-7,9-dihydro 8H-purin-8-one 93e (1.00 g, 2.03 mmol) was dissolved in 1,4-dioxane (20 mL), and 4 amino-2-fluoro-5-methylbenzamide intermediate 2 (1.36 g, 8.11 mmol), cesium carbonate (0.99 g, 3.04 mmol) and BrettPhos-G3-Pd (0.18 g, 0.20 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen atmosphere for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 80/1) to obtain the title compound 4-((9-(trans-3 ((tert-butyldiphenylsilyl)oxy)cyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide 93f (pale yellow solid, 900 mg, 71.03% yield). H NMR (400 MHz, DMSO-d6) 6 8.42 (s, 1H), 8.16 (s, 1H), 7.88 (d, 1H), 7.62 (dd, 4H), 7.55 (d, 1H), 7.49-7.40 (m, 7H), 7.24-7.16 (m, 1H), 5.13 (p, 1H), 4.76 (tt, 1H), 3.28 (s, 3H), 3.11-3.02 (m, 2H), 2.47-2.40 (m, 2H), 2.26 (s, 3H), 1.03 (s, 9H). LC-MS m/z(ESI) = 625.30 [M+1]
Step 7: 2-Fluoro-4-((9-(trans-3-hydroxycyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 93)
4-((9-(trans-3-((tert-Butyldiphenylsilyl)oxy)cyclobutyl)-7-methy-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide 93f (0.50 g, 0.80 mmol) was dissolved in 4 M hydrogen chloride-1,4-dioxane (10 mL). The reaction mixture was stirred at room temperature. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate 1,4-dioxane solution and then the pH was adjusted to 9-10 with 2 N aqueous NaOH solution, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with dichloromethane/methanol (v/v = 200:1) and concentrated to obtain the title compound 2-fluoro-4-((9-(trans-3-hydroxycyclobutyl)-7-methyl-8-oxo-8,9-dihydro 7H-purin-2-yl)amino)-5-methylbenzamide compound 93 (white solid, 300 mg, 97.02% yield). H NMR (400 MHz, DMSO-d6) 6 8.51 (s, 1H), 8.19 (s, 1H), 7.93 (d, 1H), 7.55 (d, 1H), 7.46 (s, 1H), 7.39 (s, 1H), 5.15 (d, 1H), 5.09 (q, 1H), 4.53-4.44 (m, 1H), 3.31 (s, 3H), 3.14-3.05 (m, 2H), 2.30 (s, 3H), 2.27-2.18 (m, 2H). LC-MS m/z(ESI)= 387.20 [M+1]
Example 94 2-Fluoro-4-((9-(cis-3-hydroxycyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 94)
0/ H2 N-' N N N <11N N >= F H
Compound 94 OH O 0
step I Step2 H Step3 N' 0CIN NH C~ cI N"N
CI N CI
OH OH 1A 94a 94b
H 0 N N N~N I O Step 4 >O Step 5 H 2N N N CI N>= . CI N N ==o~ F NN N
OH OH 94d Compound 94 OH 94c Step 1: Ethyl 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate (94a)
cis-3-Aminocyclobutane-1-ol hydrochloride (3.00 g, 34.48 mmol) was dissolved in acetonitrile (25 mL), and potassium carbonate (14.30 g, 103.45 mmol) and ethyl 2,4 dichloro-5-pyrimidinecarboxylate 1A (11.43 g, 51.72 mmol) were added with stirring at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)=2:1) to obtain the title compound ethyl 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate 94a (white solid, 3.3 g, 32.91% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.60 (s, 1H), 8.41 (d, 1H), 4.96 (s, 1H), 4.31 (q, 2H), 4.08-3.97 (m, 1H), 3.94-3.84 (m, 1H), 2.71-2.63 (m, 2H), 1.88-1.80 (m, 2H), 1.33-1.29 (m, 3H). LC-MS m/z(ESI) = 272.00 [M+1]
Step 2: 2-Chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid (94b)
Ethyl 2-chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylate 94a (3.30 g, 12.14 mmol) was dissolved in tetrahydrofuran/water (30 mL/15 mL), and lithium hydroxide monohydrate (1.53 g, 36.44 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to about 3-4 with 2 N HCl, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to obtain the title compound 2 chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid 94b (white solid, 2.44 g, 82.45% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.61 (d, 1H), 8.57 (s, 1H), 4.43-4.20 (m, 1H), 4.07 3.96 (m, 1H), 3.93-3.83 (m, 1H), 2.72-2.63 (m, 2H), 1.87-1.77 (m, 2H). LC-MS m/z(ESI)= 244.00 [M+1]
Step 3: 2-Chloro-9-(cis-3-hydroxycyclobutyl)-7,9-dihydro-8H-purin-8-one (94c)
2-Chloro-4-((cis-3-hydroxycyclobutyl)amino)pyrimidine-5-carboxylic acid 94b (2.44 g, 10.01 mmol) was dissolved in N,N-dimethylacetamide (20 mL), and triethylamine (1.39 mL, 10.01 mmol) and diphenylphosphoryl azide (2.16 mL, 10.01 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with dichloromethane. The organic layer was concentrated to obtain the title compound 2 chloro-9-(cis-3-hydroxycyclobutyl)-7,9-dihydro-8H-purin-8-one 94c (white solid, 0.90 g, 37.35% yield). H NMR (400 MHz, DMSO-d6) 6 11.59 (s, 1H), 8.12 (s, 1H), 4.31-4.20 (m, 1H), 4.00 3.92 (m, 1H), 3.80-3.54 (m, 1H), 2.83-2.73 (m, 2H), 2.58-2.51 (m, 2H). LC-MS m/z(ESI)= 241.00 [M+1]
Step 4: 2-Chloro-9-(cis-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one (94d)
2-Chloro-9-(cis-3-hydroxycyclobutyl)-7,9-dihydro-8H-purin-8-one 94c (0.70 g, 1.25 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dimethyl sulfate (0.28 mL, 1.25 mmol) and cesium carbonate (1.42 g, 4.36 mmol) were added with stirring at °C. The reaction mixture was stirred for 2 h. The reaction was s monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred and extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1:1) to obtain the title compound 2-chloro-9-(cis 3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one 94d (white solid, 0.32 g, 41.20% yield). H NMR (400 MHz, DMSO-d6) 6 8.33 (s, 1H), 5.31 (d, 1H), 4.34-4.23 (m, 1H), 4.02 3.91 (m, 1H), 3.34 (s, 3H), 2.81-2.72 (m, 2H), 2.58-2.51 (m, 2H). LC-MS m/z(ESI)= 255.00 [M+1] Step 5: 2-Fluoro-4-((9-(cis-3-hydroxycyclobutyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 94)
2-Chloro-9-(cis-3-hydroxycyclobutyl)-7-methyl-7,9-dihydro-8H-purin-8-one 94d (0.32 g, 1.26 mmol) was dissolved in 1,4-dioxane (8 mL), and 4-amino-2-fluoro-5 methylbenzamide intermediate 2 (0.53 g, 3.14 mmol), cesium carbonate (0.61 g, 1.88 mmol) and BrettPhos-G3-Pd (0.11 g, 0.13 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen atmosphere for 4 h. The reaction was monitored by TLC.
After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= /1) to obtain the title compound 2-fluoro-4-((9-(cis-3-hydroxycyclobutyl)-7-methyl 8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 94 (white solid, 170 mg, 35.01% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.43 (s, 1H), 8.17 (s, 1H), 7.93 (d, 1H), 7.55 (d, 1H), 7.46 (s, 1H), 7.38 (s, 1H), 5.20 (d, 1H), 4.24 (tt, 1H), 4.02-3.90 (m, 1H), 3.31 (s, 3H), 2.79 (qd, 2H), 2.60 (qd, 2H), 2.29 (s, 3H). LC-MS m/z(ESI)= 387.10 [M+1]
Example 95 2-Fluoro-4-((9-(-3-hydroxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 95)
0 HN - N N>= F2 N N
H H HO/ Compound 95 0 OH OH
Step 1 Step 2 Step 3 N NH
H HN> 1 Ok [: 0' -6 NHHCI C I AN NH H O:
95a 95b 95c 95d
0 0 H
Step 4 Step 5 Step 6 I - Step 7 CI N NH - CI 1 N NH ~ CI N N TBDPSO TBDPSO TBDPSO
95e 95f 95g
N 0
CI O Step 8 H 2N N O Step 9 H2 N N N FI NN== N H F N N N
TBDPSO TBDPSO H HO/ 951 Compound 95 95h
Step 1: tert-Butyl (3-hydroxycyclohexyl)carbamate (95b)
tert-Butyl (3-oxocyclohexyl)carbamate 95a (8.20 g, 38.45 mmol) was dissolved in tetrahydrofuran (40 mL), and sodium borohydride (4.36 g, 115.35 mmol) was added with stirring at 0 °C. The reaction mixture was stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, saturated sodium carbonate solution was slowly added to the reaction mixture, and the resulting mixture was stirred at room temperature for 3 h. Ethyl acetate was added for extraction, and the organic layer was concentrated to give the title compound tert-butyl (3 hydroxycyclohexyl)carbamate 95b (yellow liquid, 8.00 g, 96.95% yield). LC-MS m/z(ESI) = 116.10 [M+1]
Step 2: 3-Aminocyclohexan-1-ol hydrochloride (95c)
tert-Butyl (3-hydroxycyclohexyl)carbamate 95b (8.00 g, 37.16 mmol) was dissolved in 2 M hydrogen chloride-ethyl acetate (35 mL). The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate the solvent to obtain the title compound 3-aminocyclohexan-1-ol hydrochloride 95c (yellow liquid, 4.00 g, 71.05% yield). LC-MS m/z(ESI) = 116.10 [M+1]
Step 3: Ethyl 2-chloro-4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate (95d)
3-Aminocyclohexan-1-ol hydrochloride 95c (4.00 g, 26.38 mmol) was dissolved in acetonitrile (30 mL), and ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1A (8.75 g, 39.57 mmol) and potassium carbonate (10.94 g, 79.14 mmol) were added with stirring at 0 °C. The reaction mixture was warmed to room temperature and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 2:1) to obtain the title compound ethyl 2-chloro 4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate 95d (pale yellow solid, 1.40 g, 17.71% yield). LC-MS m/z(ESI)= 300.10 [M+1]
Step 4: Ethyl 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5 carboxylate (95e)
Ethyl2-chloro-4-((3-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate95d(1.10g, 3.67 mmol) was dissolved in dichloromethane (20 mL), and tert butyldiphenylchlorosilane (1.43 mL, 5.50 mmol) and imidazole (0.62 g, 9.17 mmol) were added, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 8:1) to obtain the title compound ethyl 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5 carboxylate 95e (colorless liquid, 1.90 g, 96.21% yield). LC-MS m/z(ESI) = 538.20 [M+1]
Step 5: 4-((3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5 carboxylic acid (95f)
Ethyl 4-((3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5 carboxylate 95e (1.90 g, 3.53 mmol) was dissolved in tetrahydrofuran/water (20 mL/20 mL), and lithium hydroxide monohydrate (0.44 g, 10.59 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to 3-4 with 2 N HCl, and water was added and the organics were extracted two times with ethyl acetate. The organic layer was concentrated to yield the title compound 4-((3-((tert butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid 95f (white solid, 1.10 g, 61.08% yield). LC-MS m/z(ESI)= 510.20 [M+1]
Step 6: 9-(3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9-dihydro-8H-purin-8-one (95g)
4-((3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)amino)-2-chloropyrimidine-5 carboxylic acid 95f (1.10 g, 2.16 mmol) was dissolved in N,N-dimethylacetamide (23 mL), and triethylamine (0.30 mL, 2.16 mmol) and diphenylphosphoryl azide (0.46 mL, 2.16 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1) to obtain the title compound 9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9 dihydro-8H-purin-8-one 95g (white solid, 0.31 g, 28.35% yield). LC-MS m/z(ESI)= 507.20 [M+1]
Step 7: 9-(3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7-methyl-7,9-dihydro-8H purin-8-one (95h)
9-(3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7,9-dihydro-8H-purin-8-one g (0.30 g, 0.59 mmol) was dissolved in N,N-dimethylformamide (10 mL), and dimethyl sulfate (0.06 mL, 0.59 mmol) and cesium carbonate (0.39 g, 1.18 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water. The mixture was stirred and extracted with ethyl acetate. The organic layer was concentrated to yield the title compound 9-(3-((tert butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7-methyl-7,9-dihydro-8H-purin-8-one h (colorless liquid, 0.30 g, 97.31% yield). LC-MS m/z(ESI)= 521.20 [M+1]
Step 8: 4-((9-(3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-2-fluoro-5-methylbenzamide (95i)
9-(3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)-2-chloro-7-methyl-7,9-dihydro-8H purin-8-one 95h (1.00 g, 1.92 mmol) was dissolved in 1,4-dioxane (15 mL), and 4 amino-2-fluoro-5-methylbenzamide (1.29 g, 7.68 mmol), cesium carbonate (0.94 g, 2.88 mmol) and BrettPhos-G3-Pd (0.17 g, 0.19 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen atmosphere for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to obtain the title compound 4-((9-(3-((tert-butyldiphenylsilyl)oxy)cyclohexyl) 7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide 95i (yellow solid, crude, 707 mg, 56.44% yield). LC-MS m/z(ESI)= 653.40 [M+1]
Step 9: 2-Fluoro-4-((9-(-3-hydroxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide (compound 95)
4-((9-(3-((tert-Butyldiphenylsilyl)oxy)cyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-2-fluoro-5-methylbenzamide 95i (0.65 g, 0.99 mmol) was dissolved in 4 M hydrogen chloride-1,4-dioxane (10 mL). The reaction mixture was stirred at room temperature. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate 1,4-dioxane solution and then the pH was adjusted to 9-10 with 2 N aqueous NaOH solution, and ethyl acetate was added for extraction. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = /1), followed by concentration to obtain the title compound 2-fluoro-4-((9-(-3 hydroxycyclohexyl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-5 methylbenzamide compound 95-1 (pale yellow solid, 100 mg, 24.35% yield) and compound 95-2 (white solid, 47 mg, 11.44% yield).
Compound 95-1: H NMR (400 MHz, DMSO-d) 68.48 (s, 1H), 8.18 (s, 1H), 7.98 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.41-7.35 (m, 1H), 4.79 (d, 1H), 4.22 (tt, 1H), 3.50 (tq, 1H), 3.32 (s, 3H), 2.30 (s, 3H), 2.18 (qd, 2H), 1.95-1.83 (m, 2H), 1.79 (dd, 1H), 1.68 (d, 1H), 1.40-1.26 (m, 1H), 1.24-1.15 (m, 1H). LC-MS m/z(ESI)= 415.20 [M+1] Compound 95-2: H NMR (400 MHz, DMSO-d) 68.50 (s, 1H), 8.18 (s, 1H), 7.98 (d, 1H), 7.54 (d, 1H), 7.45 (s, 1H), 7.39 (s, 1H), 4.68 (dt, 1H), 4.63 (d,1H), 4.15-4.10 (m, 1H), 3.32 (s, 3H), 2.53-2.47 (m, 1H), 2.30 (s, 3H), 2.21 (qd, 1H), 1.74 (dt, 4H), 1.57 (d, 1H), 1.42 (t, 1H). LC-MS m/z(ESI)= 415.10 [M+1]
Example 96 2-Fluoro-4-((9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide (compound 96)
H 2N N O=N NF NN N0 H
OH Intermediate 96
OH 0 N oNH 2
Step 1 Step 2 Step 3 Step 4
OH OTBDPS OTBDPS OTBDPS OTBDPS 96a 96b 96c 96d 96e
00 H NO N 0~ N~ OH N
CIN NH Step 6 CI HH Step7 CI N Step8
0 OBP DOTBDPS OTBDPS OTBDPS 96f 96g 96h
CI -O Step 9 H2N ONStep1 H2N N N H NNH 0 q OTBDPS OTBDPS OH
961 96j Compound 96 Step 1: tert-Butyl((3,4-dihydro-2H-pyran-2-yl)methoxy)diphenylsilane (96b)
(3,4-Dihydro-2H-pyran-2-yl)methanol 96a (20.0 g, 175.22 mmol) was dissolved in dichloromethane (400 mL), and tert-butyldiphenylchlorosilane (68.35 mL, 262.84 mmol) and imidazole (29.82 g, 438.06 mmol) were added, followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at room temperature overnight. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was washed with water, and the organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 100:1) to obtain the title compound tert-butyl((3,4-dihydro 2H-pyran-2-yl)methoxy)diphenylsilane 96b (colorless liquid, 42.78 g, 75.00% yield). 1H NMR (400 MHz,CDCl3):7.71-7.68 (m, 4H), 7.45-7.37 (m, 6H), 6.37- 6.35 (d, 1H), 4.68-4.65 (m, 1H), 3.96-3.93 (m, 1H), 3.83-3.77 (m, 1H), 3.71-3.67 (m, 1H), 2.08-2.00 (m, 1H), 1.99-1.92 (m, 2H), 1.74-1.71 (m, 1H), 1.02 (s, 9H). LC-MS m/z(ESI) = 353.20 [M+1]
Step 2: 6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol (96c)
tert-Butyl((3,4-dihydro-2H-pyran-2-yl)methoxy)diphenylsilane 96b (23.5 g, 66.7 mmol) was added to tetrahydrofuran (200 mL). Under nitrogen atmosphere, the reaction mixture was cooled to -78 °C, and then borane-dimethyl sulfide complex (100 mL, 2 M, 200.00 mmol) was added dropwise thereto. After addition, the reaction mixture was naturally warmed to room temperature and stirred overnight. 1 N aqueous sodium hydroxide solution was slowly added dropwise to the reaction system until no borane gas was discharged, and then 30% hydrogen peroxide (90 mL) was added to the reaction mixture. The reaction mixture was stirred at 45 °C for 2 h. Water was added and the organics were extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by reversed-phase column chromatography (acetonitrile/water (v/v) = 5:95) to obtain the title compound 6-(((tert butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol96c(colorlessliquid,18.03 g, 73.00% yield). LC-MS m/z(ESI)= 371.20 [M+1]
Step 3: 2-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3 dione (96d)
6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-ol 96c (13.0 g, 35.1 mmol) was dissolved in tetrahydrofuran (150 mL), and phthalimide (5.16 g, 35.1 mmol) and triphenylphosphine (13.8 g, 52.7 mmol) were added with stirring at room temperature, followed by the slow dropwise addition of diisopropyl azodicarboxylate (10.7 g, 52.7 mmol). The reaction mixture was heated to 70 °C and refluxed for 1 h. The reaction was s monitored by TLC. After completion of the reaction, the reaction mixture was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 100:1) to obtain the title compound 2-(6-(((tert butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3-dione 96d (white solid, 9.0 g, 51.34% yield). LC-MS m/z(ESI)= 500.20 [M+1]
Step 4: 6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-amine (96e)
2-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3 dione 96d (9.0 g, 18.0 mmol) was dissolved in methanol (50 mL), and hydrazine hydrate (1.8 g, 36.0 mmol) was added. The reaction mixture was refluxed for 2 h. The reaction was monitored by TLC. After completion of the reaction, dichloromethane (50 mL) was added, and a large amounts of solid was precipitated. Filtration and rotary evaporation drying were performed. The resulting viscos solid was washed twice with dichloromethane, and then the mixture was filtered. The organic phases were combined and concentrated under reduced pressure to yield the title compound 6-(((tert butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-amine 96e (colorless liquid, 6.3 g, 94.10% yield). LC-MS m/z(ESI)= 370.20 [M+1]
Step 5: Ethyl 4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2 chloropyrimidine-5-carboxylate (96f)
Ethyl 2,4-dichloro-5-pyrimidinecarboxylate 1A (6.94 g, 31.40 mmol) was dissolved in acetonitrile (25 mL), and potassium carbonate (8.68 g, 62.80 mmol) was added with stirring at room temperature, followed by the slow dropwise addition of a solution of 6-((tert-butyldiphenylsilyl)oxy)methyltetrahydro-2H-pyran-3-amine 96e (7.40 g, 20.93 mmol) in acetonitrile (15 mL). The reaction mixture was stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered through celite, and the filtrate was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 20:1) to obtain the title compound ethyl 4-((6 (((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2 chloropyrimidine-5-carboxylate 96f (pale yellow solid, 5.20 g, 46.86% yield). LC-MS m/z(ESI) = 554.30 [M+1]
Step 6: 4-((6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2 chloropyrimidine-5-carboxylic acid (96g)
Ethyl 4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-2 chloropyrimidine-5-carboxylate 96f (5.20 g, 9.383 mmol) was dissolved in tetrahydrofuran/water (30 mL/15 mL), and lithium hydroxide monohydrate (1.18 g, 28.15 mmol) was added. The reaction mixture was stirred at room temperature for 3 h, then heated to 50 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to 3-4 with 2 N HCl, and water and ethyl acetate were added for two extractions. The organic layer was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 4:1) to obtain the title compound 4-((6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3 yl)amino)-2-chloropyrimidine-5-carboxylic acid 96g (white solid, 2.57 g, 52.06% yield). LC-MS m/z(ESI) = 526.20 [M+1]
Step 7: 9-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9 dihydro-8H-purin-8-one (96h)
4-((6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-y)amino)-2 chloropyrimidine-5-carboxylic acid 96g (2.57 g, 4.88 mmol) was dissolved in N,N dimethylacetamide (24 mL), and triethylamine (0.68 mL, 4.88 mmol) and diphenylphosphoryl azide (1.05 mL, 4.88 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. Water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 1:1) to obtain the title compound 9-(6-(((tert butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9-dihydro-8H purin-8-one 96h (pale pink solid, 1.60 g, 62.66% yield). H NMR (400 MHz, DMSO-d6) 6 11.65 (s, 1H), 8.12 (s, 1H), 7.67-7.62 (m, 4H), 7.46
7.39 (m, 6H), 4.32-4.24 (m, 1H), 4.13-4.06 (m, 1H), 4.00-3.93 (m, 1H), 3.91-3.81 (m, 2H), 3.56 (dd, 1H), 2.53-2.47 (m, 1H), 2.07-1.98 (m, 1H), 1.85-1.72 (m, 2H), 1.01 (s, 9H). LC-MS m/z(ESI) = 523.20 [M+1]
Step 8: 9-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7 methyl-7,9-dihydro-8H-purin-8-one(96i)
9-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7,9 dihydro-8H-purin-8-one 96h (1.60 g, 3.06 mmol) was dissolved in NN dimethylformamide (15 mL), and dimethyl sulfate (0.29 mL, 3.06 mmol) and cesium carbonate (2.00 g, 6.13 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 3:2) to obtain the title compound 9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2 chloro-7-methyl-7,9-dihydro-8H-purin-8-one 96i (pale pink liquid, 1.6 g, 97.21% yield). H NMR (400 MHz, DMSO-d6) 6 8.35 (s, 1H), 7.67-7.62 (m, 4H), 7.49-7.39 (m, 6H), 4.36-4.28 (m, 1H), 4.14-4.07 (m, 1H), 4.00-3.93 (m, 1H), 3.90-3.81 (m, 2H), 3.57 (dd, 1H), 3.35 (s, 3H), 2.55-2.44 (m, 1H), 2.09-1.98 (m, 1H), 1.87-1.73 (m, 2H), 1.01 (s, 9H). LC-MS m/z(ESI)= 537.30 [M+1]
Step 9: 4-((9-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide (96j)
9-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-2-chloro-7 methyl-7,9-dihydro-8H-purin-8-one 96i (1.00 g, 1.86 mmol) was dissolved in 1,4 dioxane (20 mL), and 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (1.25 g, 7.45 mmol), cesium carbonate (0.91 g, 2.79 mmol) and BrettPhos-G3-Pd (0.17 g, 0.19 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen atmosphere for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 80/1) to obtain the title compound 4-((9-(6-(((tert-butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide 96j (yellow solid, 750 mg, 60.23% yield). LC-MS m/z(ESI)= 669.40 [M+1]
Step 10: 2-Fluoro-4-((9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide (compound 96)
4-((9-(6-(((tert-Butyldiphenylsilyl)oxy)methyl)tetrahydro-2H-pyran-3-yl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide 96j (0.50 g, 0.75 mmol) was dissolved in 4 M hydrogen chloride-1,4-dioxane (8 mL). The reaction mixture was stirred at room temperature. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate 1,4 dioxane solution and then the pH was adjusted to 9-10 with 2 N aqueous NaOH solution, and ethyl acetate was added for extraction. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1), followed by concentration to yield the title compound 2-fluoro-4-((9-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-7-methyl-8 oxo-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 96 (white solid, 170 mg, 52.83% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.55 (s, 1H), 8.18 (s, 1H), 7.76 (d, 1H), 7.55 (d, 1H), 7.49 (s, 1H), 7.39 (s, 1H), 4.63-4.58 (m, 1H), 4.30 (tt, 1H), 4.15 (t, 1H), 3.71- 3.64 (m, 2H), 3.56 -3.45 (m, 2H), 3.32 (s, 3H), 2.65-2.35 (m, 1H), 2.28 (s, 3H), 1.86 (d, 1H), 1.79-1.67 (m, 2H). LC-MS m/z(ESI) = 431.20 [M+1]
Example 97 2-Fluoro-4-((9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide(compound97)
0
HFN N N H OH
Compound 97 0 HO NHOH OH SN OH S Step1 H Se Step 6 H HN__ CI N NH HN
OH OH 97a 97b 97c 97d
H /0
Step 1:Step5 I >iO Ste6 HN N NN" Sey 6 CI-am N N ]p -c xl 9N N g,_1._mol was d 6F N N N H
OH OH OH
97e 97f Compound 97
Step 1: (6-Aminospiro[3.3]heptan-2-yl)methanol (97b)
Methyl 6-aminospiro[3.3]heptan-2-carboxylate 97a (2.0 g, 11.8 mmol) was dissolved in tetrahydrofuran (15 mL). The mixture was cooled to 0 °C, and 1 N lithium aluminum hydride solution (23.6 mL, 1 mol/L in tetrahydrofuran) was slowly added dropwise. After addition, the reaction mixture was slowly warmed to room temperature and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with methanol and concentrated under reduced pressure to evaporate the solvent. The residue was slurried with acetonitrile (50 mL), and the slurry was filtered. The filtrate was concentrated by rotary evaporation under reduced pressure to give the title compound (6-aminospiro[3.3]heptan-2-yl)methanol 97b (white solid, 1.3 g, 77.8% yield). LC-MS m/z(ESI) = 142.10 [M+1]
Step 2: Ethyl 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5 carboxylate (97c)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (2.03 g, 9.19 mmol), (6 aminospiro[3.3]heptan-2-yl)methanol 97b (1.3 g, 9.19 mmol) and potassium carbonate (1.27 g, 9.19 mmol) were dissolved in acetonitrile (25 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC. After completion of the reaction, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (v/v) = 1:1) to obtain the title compound ethyl 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2 yl)amino)pyrimidine-5-carboxylate 97c (white solid, 1.56 g, 52.2% yield). H NMR (400 MHz, Chloroform-d) 6 8.65 (s, 1H), 8.54 (d, 1H), 4.59-4.48 (m, 1H), 4.35 (q, 2H), 3.59 (dd, 2H), 2.66-2.58 (m, 1H), 2.51-2.38 (m, 2H), 2.27-2.19 (m, 1H), 2.11-2.04 (m, 2H), 2.03-1.91 (m, 2H), 1.91-1.85 (m, 1H), 1.84-1.76 (m, 1H), 1.39 (t, 3H). LC-MS m/z(ESI) = 326.10 [M+1]
Step 3: 2-Chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid (97d)
Ethyl 2-chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5 carboxylate 97c (1.07 g, 3.28 mmol) was dissolved in tetrahydrofuran/water (10 mL/10 mL), and lithium hydroxide monohydrate (0.41 g, 9.85 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate tetrahydrofuran and the pH was adjusted to about 3-4 with 2 N HCl, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with water and petroleum ether/ethyl acetate (v/v = 10/1) to obtain the title compound 2 chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid 97d (white solid, 918 mg, 93.88% yield). 'H NMR (400 MHz, DMSO-d6) 6 12.69 (s, 1H), 9.67 (d, 1H), 8.35 (s,1H), 4.42-4.31 (m, 1H), 3.30 (s, 1H), 2.55-2.46 (m, 3H), 2.40-2.31 (m, 1H), 2.28-2.19 (m, 1H), 2.10 2.00 (m, 2H), 2.00-1.91 (m, 2H), 1.83-1.69 (m, 2H). LC-MS m/z(ESI)= 298.10 [M+1]
Step 4: 2-Chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one
(97e)
2-Chloro-4-((6-(hydroxymethyl)spiro[3.3]heptan-2-yl)amino)pyrimidine-5-carboxylic acid 97d (0.91 g, 3.06 mmol) was dissolved in N,N-dimethylacetamide (12 mL), and triethylamine (0.42 mL, 3.06 mmol) and diphenylphosphoryl azide (0.66 mL, 10.01 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2 h, then heated to 110 °C and refluxed for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20:1) to obtain the title compound 2-chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro 8H-purin-8-one 97e (white solid, 516 mg, 57.28% yield). H NMR (400 MHz, DMSO-d6) 6 11.60 (s, 1H), 8.11 (s, 1H), 4.68-4.57 (m, 1H), 4.46 (t, 1H), 3.37-3.32 (m, 2H), 2.95-2.81 (m, 2H), 2.42-2.35 (m, 1H), 2.32-2.19 (m, 2H), 2.19-2.12 (m, 1H), 2.06-1.98 (m, 1H), 1.89-1.77 (m, 2H). LC-MS m/z(ESI)= 295.10 [M+1]
Step 5: 2-Chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H purin-8-one (97f)
2-Chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7,9-dihydro-8H-purin-8-one 97e (516 mg, 1.75 mmol) was dissolved in N,N-dimethylformamide (6 mL), and dimethyl sulfate (0.17 mL, 1.75 mmol) and cesium carbonate (1.14 g, 3.50 mmol) were added with stirring at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was slowly added dropwise to ice water, and the resulting mixture was stirred and extracted with ethyl acetate. The organic layer was concentrated to give the title compound 2-chloro 9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 97f (white solid, 470 mg, 86.95% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.33 (s, 1H), 4.71-4.60 (m, 1H), 4.10-3.96 (m, 2H), 3.35 (s, 1H), 3.33 (s, 3H), 2.93-2.81 (m, 2H), 2.44-2.36 (m, 1H), 2.32-2.21 (m, 2H), 2.20-2.13 (m, 1H), 2.07-1.99 (m, 1H), 1.89-1.78 (m, 2H). LC-MS m/z(ESI) = 309.10 [M+1]
Step 6: 2-Fluoro-4-((9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-8-oxo-8,9 dihydro-7H-purin-2-yl)amino)-5-methylbenzamide (compound 97)
2-Chloro-9-(6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-7,9-dihydro-8H purin-8-one 97f (100 mg, 0.32 mmol) was dissolved in 1,4-dioxane (5 mL), and 4 amino-2-fluoro-5-methylbenzamide intermediate 2 (218 mg, 1.30 mmol), cesium carbonate (158 mg, 0.49 mmol) and BrettPhos-G3-Pd (29 mg, 0.032 mmol) were added with stirring at room temperature, followed by nitrogen purging. The reaction mixture was heated to 110 °C and refluxed under nitrogen atmosphere for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1) to obtain the title compound 2-fluoro-4-((9 (6-(hydroxymethyl)spiro[3.3]heptan-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide compound 97 (white solid, 50 mg, 35.05% yield).
H NMR (400 MHz, DMSO-d6) 6 8.66 (s, 1H), 8.18 (s, 1H), 7.83 (d, 1H), 7.56 (d, 1H), 7.49 (s, 1H), 7.40 (s, 1H), 4.70-4.60 (m, 1H), 3.32 (s, 1H), 3.30 (s, 3H), 2.96-2.84 (m, 2H), 2.38-2.31 (m, 1H), 2.30-2.23 (m, 4H), 2.23-2.18 (m, 1H), 2.17-2.11 (m, 1H), 1.98 -1.91 (m, 1H), 1.88 -1.81 (m, 1H), 1.77-1.71 (m, 1H). LC-MS m/z(ESI)= 441.20 [M+1]
Example 98 4-[(9-(5-Hydroxy-octahydropentalen-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]-3-methylbenzonitrile (compound 98)
NC N N)--O NN N H
Compound 98 OH 0 0
NOH H2N N O OH
Step Step2 Step 3 N / NH Step 4 ' N NH Stp Sep2C1 i>-N 1C N
0 OH
98a 98b 98c 98d OH 98e OH
NH" NC_(: N I >=O >O ." N>==O Step 6 CI N N Step 7 N O Step 5 C N O
98f OH 98g OH Compound 98 OH Step 1: (5Z)-5-(hydroxyimino)octahydropentalen-2-one (98b)
Hydroxylamine hydrochloride (9.05 g, 130.28 mmol) and potassium carbonate (18.01, 130.28 mmol) were dissolved in 100 mL of water, and the resulting solution was added to a solution of octahydropentalene-2,5-dione 98a (20 g, 144.76 mmol) in ethanol (200 mL). The reaction mixture was refluxed at 90 °C for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove ethanol, then diluted with 100 mL of water and extracted with ethyl acetate. The organic phases were combined, dried and filtered, and the filtrate was concentrated to get a crude product, which was then purified by column chromatography (dichloromethane:methanol = 100:1) to give the title compound 5 (hydroxyimino)hexahydropentalen-2(1H)-one 98b (white solid, 7 g, 31.57% yield). 'H NMR (400 MHz, DMSO-d6) 6 10.35 (s, 1H), 2.79-2.69 (m, 2H), 2.66-2.54 (m, 2H), 2.42-2.34 (m, 2H), 2.23-2.13 (m, 2H), 2.05-1.99 (m, 1H), 1.96-1.90 (m, 1H). LC-MS m/z(ESI) = 154.10 [M+1]
Step 2: -Amino-octahydropentalen-2-ol (98c)
-(hydroxyimino)hexahydropentalen-2(1H)-one 98b (4 g, 26.11 mmol) was dissolved in tetrahydrofuran (100 mL), and cobalt chloride (12.42 g, 52.22 mmol) was added. The solution was cooled to -30 °C, and sodium borohydride (4.94 g, 130.55 mmol) was added in batches. The reaction mixture was stirred for 1 h with the temperature maintained at -25 °C to -30 °C throughout the reaction. The reaction was monitored by TLC. After completion of the reaction, saturated brine was added to the reaction mixture until no bubbles were formed. The reaction mixture was warmed to room temperature, stirred for another 0.5 h and filtered, and the filtrate was concentrated and dried to give the crude product 5-amino-octahydropentalen-2-ol 98c (oily solid, 1 g, 27.12% yield). LC-MS m/z(ESI) = 142.10 [M+1]
Step 3: Ethyl 2-chloro-4-[(5-hydroxy-octahydropentalen-2-yl)amino]pyrimidine-5 carboxylate (98d)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (2.0 g, 9.05 mmol) and 5-amino octahydropentalen-2-ol 98c (1 g, 7.08 mmol) were dissolved in acetonitrile (40 mL), and potassium carbonate (2.5 g, 18.10 mmol) was added in an ice bath. The reaction mixture was stirred at room temperature for 12 h. The reaction was monitored by TLC. After completion of the reaction, 50 mL water was added and the organics were extracted with ethyl acetate. The organic phases were combined, dried and concentrated, and the residue was purified by column chromatography (petroleum ether: acetate = 4:1) to obtain the title compound ethyl 2-chloro-4-[(5-hydroxy octahydropentalen-2-yl)amino]pyrimidine-5-carboxylate 98d (yellow solid, 1 g, 33.92% yield). H NMR (400 MHz, Chloroform-d) 6 8.63 (s, 1H), 4.48-4.41 (m, 1H), 4.33 (q, 2H), 2.58 -2.46 (m, 2H), 2.42-2.36 (m, 2H), 2.11-2.05 (m, 2H), 1.57-1.48 (m, 4H), 1.37 (t, 3H). LC-MS m/z(ESI) = 326.10 [M+1]
Step 4: 2-Chloro-4-[(5-hydroxy-octahydropentalen-2-yl)amino]pyrimidine-5-carboxylic acid (98e)
Ethyl 2-chloro-4-[(5-hydroxy-octahydropentalen-2-yl)amino]pyrimidine-5 carboxylate 98d (1 g, 3.07 mmol) was dissolved in 10 mL of tetrahydrofuran and 10 mL of water, and lithium hydroxide (0.15 g, 6.14 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove tetrahydrofuran and the pH was adjusted to 5 with 6 N hydrochloric acid, and a solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether and collected to obtain the title compound 2-chloro-4-[(5-hydroxy octahydropentalen-2-yl)amino]pyrimidine-5-carboxylic acid 98e (white solid, 0.57 g, 62.36% yield).
H NMR (400 MHz, DMSO-d6) 6 13.77 (s, 1H), 8.59 (d, 1H), 8.56 (s, 1H), 4.55 (s, 1H), 4.29-4.22 (m, 1H), 4.12-4.10 (m, 1H), 2.41-2.37 (m, 2H), 2.28-2.22 (m, 2H), 1.89 1.83 (m, 2H), 1.53-1.42 (m, 4H). LC-MS m/z(ESI)= 298.10 [M+1]
Step 5: 2-Chloro-9-(5-hydroxy-octahydropentalen-2-yl)-8,9-dihydro-7H-purin-8-one (98f)
2-Chloro-4-[(5-hydroxy-octahydropentalen-2-yl)amino]pyrimidine-5-carboxylic acid 98e (0.57 g, 1.91 mmol) was dissolved in N,N-dimethylacetamide (20 mL), and diphenyl azophosphate (0.74 g, 1.91 mmol) and triethylamine (0.19 g, 1.91 mmol) were added in an ice bath. The reaction mixture was stirred at room temperature for 1 h, heated to 90 °C and stirred for another 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was naturally cooled to room temperature, diluted with 40 mL of water and extracted with ethyl acetate. The organic phases were combined, dried and concentrated to obtain the title compound 2-chloro 9-(5-hydroxy-octahydropentalen-2-yl)-8,9-dihydro-7H-purin-8-one 98f (yellow solid, 0.5 g, 54.65% yield). H NMR (400 MHz, DMSO-d6) 6 8.11 (s, 1H), 4.67 (d, 1H), 4.58-4.49 (m, 1H), 4.05 (m, 1H), 2.40-2.24 (m, 4H), 2.06-2.01 (m, 4H), 1.39-1.33 (m, 2H). LC-MS m/z(ESI)= 295.10 [M+1]
Step 6: 2-Chloro-9-(5-hydroxy-octahydropentalen-2-yl)-7-methyl-8,9-dihydro-7H-purin-8 one (98g)
2-Chloro-9-(5-hydroxy-octahydropentalen-2-yl)-8,9-dihydro-7H-purin-8-one 98f (0.3 g, 1.02 mmol) was dissolved in dimethylformamide (10 mL), and cesium carbonate (0.66 g, 2.04 mmol) and dimethyl sulfate (0.13 g, 1.02 mmol) were added at 0 °C. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, 20 mL of water was added, and a solid was precipitated. The solid was collected by filtration and dried to give the title compound 2-chloro-9-(5-hydroxy-octahydropentalen-2-yl)-7-methyl-8,9-dihydro-7H-purin-8 one 98g (pale yellow solid, 0.14 g, 44.45% yield). H NMR (400 MHz, DMSO-d6) 6 8.34 (s, 1H), 4.63 (d, 1H), 4.60-4.52 (m, 1H), 4.10 4.01 (m, 1H), 3.34 (s, 3H), 2.40-2.24 (m, 4H), 2.07-1.99 (m, 4H), 1.40-1.33 (m, 2H). LC-MS m/z(ESI) = 309.10 [M+1]
Step 7: 4-[(9-(5-Hydroxy-octahydropentalen-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino]-3-methylbenzonitrile (compound 98)
4-Amino-3-methylbenzonitrile 5a (50 mg, 0.38 mmol), 2-chloro-9-(5-hydroxy octahydropentalen-2-yl)-7-methyl-8,9-dihydro-7H-purin-8-one 98g (117 mg, 0.38 mmol), cesium carbonate (248 mg, 0.76 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (34.45 mg, 0.04 mmol) were dissolved in dioxane (5 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100/1) to obtain the title compound 4-[(9-(5 hydroxy-octahydropentalen-2-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino] 3-methylbenzonitrile compound 98 (white solid, 60 mg, 38.12% yield). 1H NMR (400 MHz, DMSO-d6) 68.50 (s, 1H), 8.27 (d, 1H), 8.20 (s, 1H), 7.73 (dd, 1H),7.62 (d, 1H), 4.82 (d, 1H), 4.63-4.55 (in, 1H), 4.13-4.04 (in, 1H), 3.34 (s, 3H), 2.33-2.29 (in, 7H), 2.09-2.02 (in, 4H), 1.34-1.30 (in, 2H). LC-MS m/z(ESI) = 405.20 [M+1]
Example 99 2-Fluoro-4-((9-((3aS,6aS)-hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-8-oxo 8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide(compound99)
0/ H 2N N N F H
0 Compound 99 O. HOOH HOO 'O Step 1 0 Step 2 HO OH Step 3 HOV OH Step 4
0 Bn' Bn Bn- 'Boc BnBoc
99a 99b 99c 99d 99e O 0 0 N O N OH CI 0 NH Step 8 SteN Step 5 Step 6 Step 7 C CI NH Step 9
Bn'H NH 2 O 0 99f 99g 99h 99!
H 0
CI N N >==o Step 10 CI N 0 HN Kiii' N Step 11 F N N N H
0 0 99j 99k Compound 99 Step 1: Dimethyl 4-(benzylamino)cyclopentane-1,2-dicarboxylate (99b)
Dimethyl 4-oxocyclopentane-1,2-dicarboxylate 99a (5 g, 24.98 mmol) was dissolved in methanol (50 mL), and benzylamine (5.4 g, 49.96 mmol) was added, followed by the addition of drops of acetic acid. The reaction mixture was stirred at 40 °C for 0.5 h. After an intermediate was detected, sodium triacetyl borohydride (15.88 g, 74.94 mmol) was added at low temperature. The reaction mixture was stirred for another hour. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated and extracted with water and dichloromethane. The organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by column chromatography to give the title compound dimethyl 4-(benzylamino)cyclopentane-1,2-dicarboxylate 99b (white solid, 6.0 g, 82.44% yield). 'H NMR (400 MHz, CDCl3) 6 7.24 -7.15 (m, 5H), 3.68 (s, 2H), 3.62 (s, 6H), 3.37-3.27 (m, 2H), 3.13-3.06 (m, 1H), 2.20 (dd, 1H), 1.97 (t, 1H), 1.86 (dd, 1H), 1.77-1.70 (m, 1H). LC-MS m/z(ESI)= 292.10 [M+1]
Step 2: (4-(Benzylamino)cyclopentane-1,2-diyl)dimethanol (99c)
Dimethyl 4-(benzylamino)cyclopentane-1,2-dicarboxylate 99b (2.0 g, 6.86 mmol) was dissolved in tetrahydrofuran (40 mL), and lithium aluminum hydride (0.63 g, 17.15 mmol) was added in batches at 0 °C. The reaction was monitored by TLC. After completion of the reaction, the lithium aluminum hydride was quenched with 10% aqueous sodium hydroxide, and the reaction mixture was filtered. The filtrate was directly concentrated to obtain the title compound (4-(benzylamino)cyclopentane-1,2 diyl)dimethanol 99c (white solid, 1.1 g, 68.32% yield). LC-MS m/z(ESI) = 236.20 [M+1]
Step 3: tert-Butyl benzyl(3,4-bis(hydroxymethyl)cyclopentyl)carbamate (99d)
(4-(Benzylamino)cyclopentane-1,2-diyl)dimethanol 99c (8 g, 34.0 mmol) was dissolved in methanol (50 mL), and triethylamine (3.64 g, 35.97 mmol) was added, followed by the addition of di-tert-butyl dicarbonate (8.16 g, 37.40 mmol). The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 50:1) to obtain the title compound tert-butyl benzyl tert-butyl benzyl(3,4 bis(hydroxymethyl)cyclopentyl)carbamate 99d (white solid, 7.18 g, 63% yield). 'H NMR (400 MHz, Chloroform-d) 6 7.32-7.16 (m, 5H), 4.37 (s, 2H), 3.72-3.61 (m,1H), 3.38-3.32 (m, 2H), 1.90-1.75 (m, 4H), 1.47-1.44 (m, 3H), 1.38 (s, 9H). LC-MS m/z(ESI) = 236.10 [M+1]
Step 4: tert-Butyl benzyl (hexahydro-1H-cyclopenta[c]furan-5-yl)carbamate (99e)
tert-Butyl benzyl (3,4-bis(hydroxymethyl)cyclopentyl)carbamate 99d (8 g, 23.85 mmol) was dissolved in tetrahydrofuran (100 mL), and n-butyllithium (10.7 mL, 26.75 mmol, 2.5 mol/L) was added dropwise at -20 °C under nitrogen atmosphere. The reaction mixture was stirred for 1 h, and a solution ofp-toluenesulfonyl chloride (5 g, 26.23 mmol) in tetrahydrofuran (10 mL) was added dropwise. The reaction mixture was stirred for 0.5 h, and n-butyllithium (10.7 mL, 26.75 mmol, 2.5 mol/L) was added dropwise again. The reaction mixture was stirred for 10 min and heated at reflux for 48 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with ice water and extracted three times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 5:1) to obtain the title compound tert-butyl benzyl(hexahydro-1H cyclopenta[c]furan-5-yl)carbamate 99e (white solid, 4.0 g, 52.84% yield). 'H NMR (400 MHz, CDCl3) 6 7.32-7.17 (m, 5H), 4.37 (s, 2H), 3.73-3.63 (m, 2H), 3.40-3.34 (m, 2H), 2.06-1.25 (m, 16H). LC-MS m/z(ESI) = 262.10 [M+1]
Step 5: N-Benzylhexahydro-1H-cyclopenta[c]furan-5-amine (99f)
tert-Butyl benzyl(hexahydro-1H-cyclopenta[c]furan-5-yl)carbamate 99e (4.0 g, 12.60 mmol) was dissolved in dichloromethane (10 mL), and a solvent mixture of dichloromethane/trifluoroacetic acid (v/v = 1:1) was added. The reaction mixture was stirred at room temperature for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with water, and the aqueous phase was adjusted to be slightly basic with sodium bicarbonate. Dichloromethane was added for three extractions, and the organic phase was dried over anhydrous sodium sulfate and concentrated to give the title compound N benzylhexahydro-1H-cyclopenta[c]furan-5-amine 99f (white solid, 1.1 g, 40.1% yield). LC-MS m/z(ESI) = 218.10 [M+1]
Step 6: Hexahydro-1H-cyclopenta[c]furan-5-amine (99g)
N-benzylhexahydro-1H-cyclopenta[c]furan-5-amine 99f (1.2 g, 5.52 mmol) was dissolved in ethanol (35 mL), and then 2 drops of acetic acid were added, followed by the addition of palladium hydroxide (1.2 g, 8.54 mmol). The system was purged with hydrogen, and the reaction mixture was stirred at room temperature for 5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by column chromatography to yield the title compound hexahydro-1H-cyclopenta[c]furan-5-amine 99g (white solid, 613 mg, 87.6% yield). 'H NMR (400 MHz, DMSO-d6) 6 4.01-4.06 (m, 1H), 3.72 (t, 2H), 3.13-3.48 (m, 4H), 2.44-2.48 (m, 1H), 1.96-2.17 (m, 2H), 1.97-1.72 (m, 1H), 1.23-1.37 (m, 1H). LC-MS m/z(ESI) = 128.10 [M+1]
Step 7: Ethyl 2-chloro-4-((hexahydro-1H-cyclopenta[c]furan-5-yl)amino)pyrimidine-5 carboxylate (99h)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (435 mg, 1.97 mmol) and potassium carbonate (816.8 mg, 5.91 mmol) were dissolved in acetonitrile (20 mL), and hexahydro-1H-cyclopenta[c]furan-5-amine 99g (300 mg, 2.36 mmol) was added at °C. The reaction mixture was warmed to room temperature and stirred for 20 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted three times with ethyl acetate. The organic layer was washed once with saturated brine, dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 10:1) to obtain the title compound ethyl 2-chloro
4-((hexahydro-1H-cyclopenta[c]furan-5-yl)amino)pyrimidine-5-carboxylate 99h (white solid, 413 mg, 67.7% yield). LC-MS m/z(ESI) = 312.10 [M+1]
Step 8: 2-Chloro-4-((hexahydro-1H-cyclopenta[c]furan-5-yl)amino)pyrimidine-5-carboxylic acid (99i)
Ethyl 2-chloro-4-((hexahydro-1H-cyclopenta[c]furan-5-yl)amino)pyrimidine-5 carboxylate 99h (680 mg, 2.18 mmol) was dissolved in 5 mL of tetrahydrofuran and 5 mL of water, and lithium hydroxide (104.38 mg, 4.36 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 4, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 2-chloro-4-((hexahydro-1H-cyclopenta[c]furan-5 yl)amino)pyrimidine-5-carboxylic acid 99i (white solid, 520 mg, 67.26% yield). LC-MS m/z(ESI)= 284.10 [M+1]
Step 9: 2-Chloro-9-(hexahydro-1H-cyclopenta[c]furan-5-yl)-7,9-dihydro-8H-purin-8-one (99j)
2-Chloro-4-((hexahydro-1H-cyclopenta[c]furan-5-yl)amino)pyrimidine-5-carboxylic acid 99i (0.5 g, 1.76 mmol) was dissolved in N,N-dimethylacetamide (20 mL), and diphenylazophosphate (0.68 g, 1.76 mmol) and triethylamine (0.18 g, 1.76 mmol) were added in an ice bath. The reaction mixture was stirred at room temperature for 1 h, then heatedto 90 °C and stirred for another 3 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was naturally cooled to room temperature, and 40 mL water was added and the organics were extracted with ethyl acetate. The organic phases were combined, dried and concentrated to yield the title compound 2-chloro-9-(hexahydro-1H-cyclopenta[c]furan-5-yl)-7,9-dihydro-8H-purin 8-one 99j (yellow solid, 0.45 g, 54.65% yield). LC-MS m/z(ESI)= 281.00 [M+1]
Step 10: 2-Chloro-9-(hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-7,9-dihydro-8H-purin 8-one (99k)
2-Chloro-9-(hexahydro-1H-cyclopenta[c]furan-5-yl)-7,9-dihydro-8H-purin-8-one 99j (0.45 g, 0.96 mmol) was dissolved in dimethylformamide (10 mL), and cesium carbonate (0.47 g, 1.44 mmol) and dimethyl sulfate (0.11 mL, 1.15 mmol) were added at 0 °C. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, then 20 mL of water was added, and a solid was precipitated. The solid was collected by filtration and dried to yield the title compound 2-chloro-9-(hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-7,9 dihydro-8H-purin-8-one 99k (pale yellow solid, 0.14 g, 49.48% yield).
H NMR (400 MHz, DMSO-d6) 6 8.35 (s, 1H), 5.49-5.21 (m, 1H), 3.82-3.77 (m, 2H), 3.37 (s, 3H), 3.35-3.27 (m, 2H), 3.23-3.18 (m, 2H), 2.96-2.84 (m, 1H), 2.15-2.03 (m, 2H), 1.94-1.85 (m, 2H). LC-MS m/z(ESI) = 295.10 [M+1]
Step 11: 2-Fluoro-4-((9-((3aS,6aS)-hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-8-oxo 8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide (compound 99)
2-Chloro-9-((3aS,6aS)-hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-7,9-dihydro 8H-purin-8-one 99k (100 mg, 0.34 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (114 mg, 0.68 mmol), cesium carbonate (221 mg, 0.68 mmol) and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1' biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (30 mg, 0.03 mmol) were dissolved in 1,4-dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 10/1) to obtain the title compound 2-fluoro-4-((9 ((3aS,6aS)-hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide compound 99 (white solid, 8 mg, 6% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.60 (s, 1H), 8.18 (s, 1H), 7.64 (d, 1H), 7.54 (d, 1H), 7.48 (s, 1H), 7.36 (s, 1H), 3.74 (t, 1H), 3.68 (t, 1H), 3.32 (s, 3H), 3.22-3.13 (m, 3H), 2.81-2.71 (m, 1H), 2.25 (s, 3H), 2.03-1.99 (m, 2H), 1.92-1.84 (m, 3H). LC-MS m/z(ESI) = 427.2 [M+1]
Example 100 4-((9-(Hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-3-methylbenzonitrile (compound 100)
NC ,,a NNI N
CN N H
Compound 100 0
CI N N N N - NH 2 H
99k 0 5a Compound 100 4-Amino-3-methylbenzonitrile 5a (50 mg, 0.38 mmol), 2-chloro-9-(hexahydro-1H cyclopenta[c]furan-5-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 99k (112 mg, 0.38 mmol), cesium carbonate (247.62 mg, 0.76 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (34.45 mg, 0.04 mmol) were dissolved in dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography
(dichloromethane/methanol (v/v) = 100/1) to obtain the title compound 4-((9 (hexahydro-1H-cyclopenta[c]furan-5-yl)-7-methyl-8-oxo--7-methyl-8-oxohydro-7H purin-2-yl)amino)-3-methylbenzonitrile compound 100 (pale yellow solid, 45 mg, 30.09% yield). 1H NMR (400 MHz, Chloroform-d) 6 8.43 (d, 1H), 7.93 (s, 1H), 7.53 (dd, 1H), 7.48 (d, 1H), 7.05 (s, 1H), 5.47-5.40 (m, 1H), 3.96 (q, 2H), 3.48 (dd, 1H), 3.43 (s, 3H), 3.37 (dd, 1H), 5.38-5.31 (m, 1H), 2.38 (s, 3H), 2.92- 2.21 (m, 5H). LC-MS m/z(ESI) = 391.20 [M+1]
Example 101 4-((9-(1,1-Dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-3-methylbenzonitrile(compound101)
NC - N N N, NN N H
0 Compound 101 0 0
N O N' OH 0te 1, Ste 7( Step 3 CN NH Step CI N NH
CI N CI
00 00 1A l1a 101b
H N N0 N NC ,a N
CI N N Step 4 C N NO Step 5 NC N N o H
60 060 0 101c 101d Compound 101 Step 1: Ethyl 2-chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5 carboxylate (101a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (8 g, 36 mmol) and potassium carbonate (9.9 g, 72 mmol) were dissolved in acetonitrile (30 mL), and 4 aminotetrahydro-2H-thiopyran-1,1-dioxide (6.6 g, 36 mmol) was added at 0 °C. The reaction mixture was warmed to room temperature and stirred for 20 h. The reaction was monitored by TLC. After completion of the reaction, 30 mL of water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration and dried to obtain the title compound ethyl 2-chloro-4-((1,1-dioxidotetrahydro-2H thiopyran-4-yl)amino)pyrimidine-5-carboxylate 101a (brown solid, 6.3 g, 76% yield). 1H NMR (400 MHz DMSO) 6 8.64 (s, 1H), 8.33 (d, 1H), 4.34-4.30 (m,3H), 3.42-3.35 (m, 1H), 3.08 (d, 1H), 2.24-2.06 (m, 4H), 1.31(t, 3H). LC-MS m/z(ESI) = 334.20 [M+1]
Step 2: 2-Chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5 carboxylic acid (101b)
Ethyl 2-chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5 carboxylate 101a (3 g, 9 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL of water, and lithium hydroxide (756 mg, 18 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to about 4, and a brown solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1), concentrated and dried to obtain the title compound 2-chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4 yl)amino)pyrimidine-5-carboxylic acid 101b (white solid, 2.3 g, 84% yield). 'H NMR (400 MHz DMSO) 6 13.80 (s, 1H), 8.60 (s, 1H), 8.57 (d,1H), 4.37-4.29 (m, 1H), 3.50-3.27 (m, 2H), 3.07 (d, 2H), 2.25-2.21 (m, 2H), 2.08-1.98 (m, 2H). LC-MS m/z(ESI) = 306.20 [M+1]
Step 3: 2-Chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7,9-dihydro-8H-purin-8-one (101c)
2-Chloro-4-((1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino)pyrimidine-5 carboxylic acid 101b (2.3 g, 7.5 mmol) was dissolved in dimethylacetamide (10 mL), and triethylamine (757 mg, 7.5 mmol) and diphenylphosphoryl azide (2 g, 7.5 mmol) were added. The reaction mixture was gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into 30 mL of water, and a brown solid was precipitated. The solid was collected by filtration to obtain the title compound 2-chloro-9-(1,1 dioxidotetrahydro-2H-thiopyran-4-yl)-7,9-dihydro-8H-purin-8-one 101c (white solid, 1.7 g, 58% yield). H NMR (400 MHz DMSO) 6 11.67 (s, 1H), 8.14 (s, 1H), 4.69-4.61 (m,1H), 3.55-3.45 (m, 2H), 3.13 (d, 2H), 2.94-2.77 (m, 2H), 2.14-2.02 (m, 2H). LC-MS m/z(ESI) = 303.20 [M+1]
Step 4: 2-Chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9-dihydro-8H purin-8-one (101d)
2-Chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7,9-dihydro-8H-purin-8-one 101c (1.7 g, 5.6 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (709 mg, 5.6 mmol) and cesium carbonate (2.7 g, 8.4 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, 10 mL of water was added, and a solid was precipitated. The solid was collected by filtration to obtain the title compound 2-chloro-9-(1,1 dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9-dihydro-8H-purin-8-one 101d (white solid, 1.3 g, 82% yield). 'H NMR (400 MHz DMSO) 6 8.33 (s, 1H), 4.26 (s, 1H), 3.33 (s, 3H), 2.99 (s, 2H), 2.11-2.04 (m, 2H), 1.80-1.61 (m, 2H), 1.44 (d, 2H).
LC-MS m/z(ESI)= 317.20 [M+1]
Step 5: 4-((9-(1,1-Dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-3-methylbenzonitrile (compound 101)
2-Chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9-dihydro-8H purin-8-one 101d (200 mg, 0.6 mmol), 4-amino-3-methylbenzonitrile (83.5 mg, 0.6 mmol), cesium carbonate (587 mg, 1.8 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (55 mg, 0.06 mmol) were dissolved in dioxane (3 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30/1) to obtain the title compound 4-((9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-8-oxo 8,9-dihydro-7H-purin-2-yl)amino)-3-methylbenzonitrile compound 101 (white solid, mg, 26% yield). 1H NMR (400 MHz DMSO) 68.57 (s, 1H), 8.32 (d, 1H), 8.23 (s, 1H), 7.62-7.60 (m, 2H), 4.68-4.62 (m, 1H), 3.52-3.45 (m, 2H), 3.18-3.15 (m, 2H), 3.00-2.89 (m, 2H), 2.34 (s,3H), 2.08 (d, 2H). LC-MS m/z(ESI) = 413.20 [M+1]
Example 102 3-Methyl-4-((7-methyl-8-oxo-9-((tetrahydrofuran-3-yl)methyl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 102)
0I H2N N N ' N<N N H
Compound 102 O 0 0 N H H Stepte I C - S5N e 1p Step3 N Eh 2N(t r NHe CItaN oCI N N 0 0
IA 102a 102b 102c
07 Step 4 >zO e5HN I: N N
CI N N -j N N N '-
102d Compound 102 Step 1: Ethyl 2-chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylate (102a)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (5 g, 22.6 mmol) and potassium carbonate (6.2 g, 44.8 mmol) were dissolved in acetonitrile (20 mL), and (tetrahydrofuran-3-yl)formamide (2.3 g, 22.6 mmol) was added at 0 °C. The reaction mixture was warmed to room temperature and stirred for 20 h. The reaction was monitored by TLC. After completion of the reaction, 30 mL of water was added to the reaction mixture, and the resulting mixture was extracted with ethyl acetate. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (n-hexane/ethyl acetate (v/v) = 10:1) to obtain the title compound ethyl 2-chloro-4-(((tetrahydrofuran-3 yl)methyl)amino)pyrimidine-5-carboxylate 102a (white solid, 4.4 g, 85% yield). H NMR (400 MHz DMSO) 6 8.58-8.61 (m, 2H), 4.31 (q, 2H), 3.78-3.73 (m,1H), 3.69 3.58 (m, 2H), 3.48-3.44 (m, 3H), 2.57-2.53 (m, 1H), 1.98-1.91 (m, 1H), 1.62-1.58 (m, 1H), 1.31 (t, 3H). LC-MS m/z(ESI) = 286.20 [M+1]
Step 2: 2-Chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylic acid (102b)
Ethyl 2-chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylate 102a (4.4 g, 15.3 mmol) was dissolved in 10 mL of tetrahydrofuran and 5 mL of water, and lithium hydroxide (736 mg, 30.6 mmol) was added. The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove tetrahydrofuran and the pH was adjusted to pH 4-5, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed twice with petroleum ether/ethyl acetate (v/v = 10/1) and concentrated to obtain the title compound 2-chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5 carboxylic acid 102b (white solid, 3.4 g, 80% yield). 'H NMR (400 MHz DMSO) 6 13.75 (s, 1H), 8.75 (s, 1H), 8.57 (s, 1H), 3.78-3.73 (m, 3H), 3.47-3.43 (m,3H), 2.58-2.52 (m, 1H), 1.98-1.89 (m, 3H), 1.63-1.55 (m, 1H). LC-MS m/z(ESI) = 259.20 [M+1]
Step 3: 2-Chloro-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one (102c)
2-Chloro-4-(((tetrahydrofuran-3-yl)methyl)amino)pyrimidine-5-carboxylic acid 102b (3.4 g, 13.0 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (1.58 g, 13 mmol) and diphenylphosphoryl azide (3.6 g, 13 mmol) were added. The reaction mixture was gradually heated to 120 °C and stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 5:1 to 1:10) to obtain the title compound 2-chloro 9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one 102c (white solid, 1.46 g, 50% yield). H NMR (400 MHz DMSO) 6 11.65 (s, 1H), 8.14 (s, 1H), 3.80-3.74 (m, 3H), 3.66-3.57 (m, 2H), 3.52-3.49 (m, 1H), 2.73-2.69 (m, 1H), 1.98-1.89 (m, 1H), 1.67-1.60 (m, 1H). LC-MS m/z(ESI)= 255.20 [M+1]
Step 4: 2-Chloro-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one (102d)
2-Chloro-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one 102c (1.6 g, 5.2 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (663 mg, 5.2 mmol) and cesium carbonate (2.4 g, 7.7 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, then 10 mL of water was added, and a solid was precipitated. The solid was collected by filtration to obtain the title compound 2-chloro-7-methyl-9 ((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one 102d (white solid, 1.2 g, % yield). H NMR (400 MHz DMSO) 6 8.35 (s, 1H), 3.81-3.75 (m, 3H), 3.65-3.57 (m, 2H), 3.52 3.49 (m, 1H), 3.37 (s, 3H), 2.73-2.70 (m, 1H), 1.96-1.91 (m, 1H), 1.67-1.61 (m, 1H). LC-MS m/z(ESI) = 268.20 [M+1]
Step 5: 3-Methyl-4-((7-methyl-8-oxo-9-((tetrahydrofuran-3-yl)methyl)-8,9-dihydro-7H purin-2-yl)amino)benzamide (compound 102)
2-Chloro-7-methyl-9-((tetrahydrofuran-3-yl)methyl)-7,9-dihydro-8H-purin-8-one 102d (300 mg, 1.12 mmol), 4-amino-3-methylbenzamide 2a (250 mg, 1.68 mmol), cesium carbonate (730 mg, 2.24 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (100 mg, 0.11 mmol) were dissolved in 1,4-dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 100 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into ice water, and the solid was collected and purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100:1 to 50:1) to obtain the title compound 3 methyl-4-((7-methyl-8-oxo-9-((tetrahydrofuran-3-yl)methyl)-8,9-dihydro-7H-purin-2 yl)amino)benzamide compound 102 (pale yellow solid, 45 mg, 10.37% yield). 'H NMR (400 MHz, DMSO-d) 6 10.21 (s, 1H), 8.50 (s, 1H), 8.10 (s, 1H), 7.83-7.64 (m, 4H), 7.16 (s, 1H), 3.78-3.74 (m, 3H), 3.69-3.59 (m, 2H), 3.55-3.52 (m, 1H), 3.32 (s, 3H), 2.81-2.71 (m, 1H), 2.29 (s, 3H), 1.96-1.88 (m, 1H), 1.69-1.61 (m, 1H). LC-MS m/z(ESI)= 383.20 [M+1].
Example 103 9-Cyclohexyl-2-((4-methoxy-2-methylphenyl)amino)-7-methyl-7,9-dihydro-8H purin-8-one (compound 103)
N N N
N )= _'[:'NN H
Compound 103
N O0 N
H CI N N N
68d Compound 103 2-Chloro-9-cyclohexyl-7-methyl-7,9-dihydro-8H-purin-8-one 68d (150 mg, 0.56 mmol), 4-methoxy-2-methylaniline (154 mg, 1.12 mmol), cesium carbonate (367 mg, 1.12 mmol) and methanesulfonato(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6' tri-i-propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (76 mg, 0.08 mmol) were dissolved in 1,4 dioxane (50 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 10/1) followed by Pre-HPLC to obtain the title compound 9-cyclohexyl-2-((4-methoxy-2-methylphenyl)amino)-7-methyl-7,9 dihydro-8H-purin-8-one compound 103 (white solid, 53 mg, 26% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.26 (s, 1H), 7.94 (s, 1H), 7.30 (d, 1H), 6.78 (d, H), 6.71 (dd, 1H), 4.134.11 (m, 1H), 3.72 (s, 3H), 3.25 (s, 3H), 2.25-2.18,2H), 2.17 (s, 3H), 1.79 (d, 2H), 1.65 (t, 3H), 1.35-1.22 (m, 2H), 1.11 (t, 1H). LC-MS m/z(ESI)= 368.20 [M+1]
Example 104 2-Fluoro-4-((9-(3-fluorotetrahydro-2H-pyran-4-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide(compound104)
0/
H2 N N N N F F )[:N<'1N N F H F
0
Compound 104 0 0 o 0 NH 2 N" O Ne OH Step I F Step 2 F Step 3 I F Step 44Stopp4 SN NH
0 0 104a 104b 104c 104d 1049
H 0 N N N N" N Step 6 O Step S N 7 )== OFOi Step 8 H 2N N bF O O H F
104f 1049 104h Compound 104 Step 1: 3-Fluorotetrahydro-4H-pyran-4-one (104b)
Tetrahydro-4H-pyran-4-one 104a (5 g, 49.94 mmol) and 1-chloromethyl-4-fluoro-1,4 diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (21.23 g, 59.93 mmol) were dissolved in acetonitrile/water (20:1 mL). The reaction mixture was heated to 80 °C and stirred for 8 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to evaporate acetonitrile, and water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)= 1:1) to obtain the title compound 3-fluorotetrahydro-4H-pyran-4-one 104b (colorless liquid, 4.4 g, 74.88% yield). LC-MS m/z(ESI)= 119.10 [M+1]
Step 2: 3-Fluorotetrahydro-2H-pyran-4-amine (104c)
3-Fluorotetrahydro-4H-pyran-4-one 104b (1.4 g, 11.85 mmol) was dissolved in absolute methanol (10 mL), and ammonium formate (7.47 g, 118.53 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 0.5 h, and then sodium cyanoborohydride (894 mg, 14.22 mmol) was added. The reaction mixture was stirred for 5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with saturated sodium bicarbonate, concentrated to dryness by rotary evaporation under reduced pressure, filtered, and washed 3 times with methanol:dichloromethane (40:1). The filtrate was concentrated to yield the title compound 3-fluorotetrahydro-2H-pyran-4-amine 104c (white solid, crude, 5.3 g). LC-MS m/z(ESI) = 120.10 [M+1]
Step 3: Ethyl 4-((3-fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)pyrimidine-5 carboxylate (104d)
Ethyl 2,4-dichloropyrimidine-5-carboxylate 1A (3.3 g, 14.20 mmol) and potassium carbonate (4.9 g, 35.50 mmol) were dissolved in acetonitrile (20 mL), and 3 fluorotetrahydro-2H-pyran-4-amine 104c (1.41 g, 11.83 mmol) was added with stirring at room temperature. The reaction mixture was heated to 30 °C and stirred for 7 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filter cake was washed with dichloromethane and ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) = 10:1) to obtain the title compound ethyl 4-((3-fluorotetrahydro-2H-pyran-4-yl)amino)-2 (methylthio)pyrimidine-5-carboxylate 104d (white solid, 1.85 g, 49.57% yield). LC-MS m/z(ESI)= 316.10 [M+1]
Step 4: 4-((3-Fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid (104e)
Ethyl 4-((3-fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)pyrimidine-5 carboxylate 104d (1.85 g, 5.87 mmol) was dissolved in 10 mL of tetrahydrofuran and mL of water, and sodium hydroxide (704 mg, 17.60 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran and the pH was adjusted to 3-4, and ethyl acetate was added for extraction. The organic phase was concentrated to yield the title compound 4-((3-fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)pyrimidine-5 carboxylic acid 104e (white solid, 1.73 g, 104.3% yield). LC-MS m/z(ESI)= 288.10 [M+1]
Step 5: 9-(3-Fluorotetrahydro-2H-pyran-4-yl)-2-(methylthio)-7,9-dihydro-8H-purin-8-one (104f)
4-((3-Fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid 104e (1.73 g, 6.02 mmol) was dissolved in dimethylacetamide (20 mL), and triethylamine (835 [L, 6.02 mmol) and diphenylphosphoryl azide (1.3 mL, 6.02 mmol) were added. The reaction mixture was stirred at room temperature for 1 h, then heated to 100 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, water was added to the reaction mixture, and a solid was precipitated. The mixture was filtered, and the filter cake was eluted with water and petroleum ether and concentrated under reduced pressure to give the title compound ethyl 4-(3 fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)-pyrimidine-5-carboxylate 104f (white solid, 1.27 g, 73.89% yield). LC-MS m/z(ESI)= 285.10 [M+1]
Step 6: 9-(3-Fluorotetrahydro-2H-pyran-4-yl)-7-methyl-2-(methylthio)-7,9-dihydro-8H purin-8-one (104g)
Ethyl 4-(3-fluorotetrahydro-2H-pyran-4-yl)amino)-2-(methylthio)-pyrimidine-5 carboxylate 104f (1.27 g, 4.45 mmol) was dissolved in dimethylformamide (10 mL), and dimethyl sulfate (433 [L, 4.45 mmol) and cesium carbonate (1.84 g, 13.35 mmol) were added at 0 °C. The reaction mixture was stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, then water was added and the organics were extracted with ethyl acetate, and the organic layer was concentrated to obtain the title compound 9-(3-fluorotetrahydro-2H-pyran-4-yl)-7-methyl-2 (methylthio)-7,9-dihydro-8H-purin-8-one 104g (white solid, 800 mg, 60.27% yield). LC-MS m/z(ESI)= 299.10 [M+1]
Step 7: 9-(3-Fluorotetrahydro-2H-pyran-4-yl)-7-methyl-2-(methylsulfonyl)-7,9-dihydro-8H purin-8-one (104h)
9-(3-Fluorotetrahydro-2H-pyran-4-yl)-7-methyl-2-(methylthio)-7,9-dihydro-8H purin-8-one 104g (800 mg, 2.68 mmol) was dissolved in dichloromethane (6 mL), and m-chloroperoxybenzoic acid (1.85 g, 10.73 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 100:1 to 60:1) to obtain the title compound 9-(3-fluorotetrahydro-2H-pyran-4 yl)-7-methyl-2-(methylsulfonyl)-7,9-dihydro-8H-purin-8-one 104h (white solid, 1.05 g, 119% yield). LC-MS m/z(ESI)= 331.10 [M+1]
Step 8: 2-Fluoro-4-((9-(3-fluorotetrahydro-2H-pyran-4-yl)-7-methyl-8-oxo-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 104)
9-(3-Fluorotetrahydro-2H-pyran-4-yl)-7-methyl-2-(methylsulfonyl)-7,9-dihydro-8H purin-8-one 104h (300 mg, 0.91 mmol), 4-amino-3-methylbenzamide intermediate 2 (458 mg, 2.72 mmol) and potassium tert-butoxide (305 mg, 2.72 mmol) were dissolved in dimethylformamide (10 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate (v/v)= 5:1 to 1:1) to get the title compound 2-fluoro-4 ((9-(3-fluorotetrahydro-2H-pyran-4-yl)-7-methyl-8-oxo-8,9-dihydro-7H-purin-2 yl)amino)-5-methylbenzamide compound 104-1 (pale yellow solid, 130 mg, 34.21% yield) and compound 104-2 (white solid, 85 mg, 22.37% yield). Compound 104-1: H NMR (400 MHz, DMSO-d6) 6 8.62 (s, 1H), 8.23 (s, 1H), 7.84 (d, 1H), 7.54 (d, 1H), 7.46 (s, 1H), 7.40 (s, 1H), 5.46-5.37 (m, 0.5H), 5.34-5.25 (m, 0.5H), 4.63-4.51 (m, 1H), 4.23-4.16 (m, 1H), 3.97-3.90 (m, 1H), 3.54-3.45 (m, 1H), 3.41-3.34 (m, 4H), 2.60-2.53 (m, 1H), 2.29 (s, 3H), 1.97-1.89 (m, 1H). LC-MS m/z(ESI)= 419.20 [M+1] Compound 104-2: H NMR (400 MHz, DMSO-d6) 6 8.50 (s, 1H), 8.22 (s, 1H), 7.92 (d, 1H), 7.54 (d, 1H), 7.45 (s, 1H), 7.39 (s, 1H), 4.82 (s, 0.5H), 4.69 (s, 0.5H), 4.67-4.61 (m, 0.5H), 4.59-4.52 (m, 0.5H), 4.09-3.98 (m, 2H), 3.75-3.70 (m, 0.5H), 3.66-3.61 (m, 0.5H), 3.60-3.52 (m, 1H), 3.49-3.37 (m, 1H), 3.35 (s, 3H), 2.29 (s, 3H), 1.81 (dd, 1H). LC-MS m/z(ESI)= 419.20 [M+1]
Example 105 4-((7-(Ethyl-d5)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 105)
H2N N N
H
0
Compound 105
0 0 N'~O N'-I OH N N O' Step 1 S Step 2 Step 3 >0 A S N" NH S N NH - S N N ~S N CIb
0 0
105a 105b 105c 105d
DDDD D D 0 DD D
Step 4 NA NtepN 1 o Step 5 NSN N )= N1 Step 6 H2N H2 AN N >==O S N>= N F N N H
0 0 0
105e 105f Compound 105 Step 1: Ethyl 2-(methylthio)-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate (105b)
Ethyl 4-chloro-2-(methylthio)pyrimidine-5-carboxylate 105a (500 g, 2.15 mol) and tetrahydro-2H-pyran-4-amine hydrochloride (300 g, 2.19 mol) were dissolved in acetonitrile (3000 mL) and water (500 mL), and after the solution was stirred several times, potassium carbonate (600 g, 4.3 mol) was added. The reaction mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was filtered, and the filter residue was washed with ethyl acetate (1000 mL). The filtrate was concentrated to get a crude product, which was then extracted with ethyl acetate and water. The organic phase was concentrated to obtain the title compound ethyl 2-(methylthio)-4-((tetrahydro-2H pyran-4-yl)amino)pyrimidine-5-carboxylate 105b (white solid, 642 g, 99.69% yield). H NMR (400 MHz DMSO) 6 8.56 (s, 1H), 8.17 (d, 1H), 4.31-4.26 (m, 2H), 4.25-4.21 (m, 2H), 3.88-3.84 (m, 2H), 3.48-3.42 (m, 2H), 2.48 (s, 3H), 1.94-1.90 (m, 2H), 1.60 1.51 (m, 2H), 1.32-1.29 (m, 3H). LC-MS m/z (ESI)= 298.10 [M+1]
Step 2: 2-(Methylthio)-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid (105c)
Ethyl 2-(methylthio)-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate 105b (640 g, 2.15 mol) was dissolved in tetrahydrofuran (1000 mL)/water (1200 mL)/methanol (200 mL), and sodium hydroxide (172.16 g, 4.30 mol) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to remove the organic solvent and the pH was adjusted to 3-4 with 12 N hydrochloric acid, and a white solid was precipitated. The mixture was filtered, and the filter cake was washed with petroleum ether and collected to yield the title compound 2 (methylthio)-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid 105c (white solid, 640 g, crude, 110.4% yield). 'H NMR (400 MHz DMSO) 6 8.52 (s, 1H), 8.36 (d, 1H), 4.22-4.18 (m,1H), 3.86-3.83 (m, 2H), 3.47-3.41 (m, 2H), 2.46 (s, 3H), 1.94-1.89 (m, 2H), 1.56-1.48 (m, 2H).
LC-MS m/z (ESI)= 270.10 [M+1]
Step 3: 2-(Methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one (105d)
2-(Methylthio)-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid 105c (630 g, 2.34 mol) was dissolved in dimethylacetamide (4000 mL), and diphenylphosphoryl azide (504 mL, 2.34 mol) was added with stirring at room temperature. The reaction mixture was stirred for 10 min, and then triethylamine (325 mL, 2.34 mol) was added. The reaction mixture was stirred for another 2 h, then heated to 120 °C and stirred for 1.5 h, during which a large amount of gas was produced. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was directly concentrated to remove most of the solvent. The residual liquid was poured into 6 L of ice water, and a large amounts of solid was precipitated. After the mixture was stirred for 0.5 h, the solid was collected by filtration and washed 3 times with water, and concentrated under reduced pressure to yield the title compound 2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro 8H-purin-8-one 105d (white solid, 880 g, crude). 'H NMR (400 MHz DMSO) 6 11.35 (s, 1H), 8.12 (s, 1H), 4.44-4.38 (m, 1H), 4.00 43.97 (m, 2H), 3.45-3.41 (m, 2H), 2.55-2.45 (m, 5H), 1.69-1.65 (m, 2H). LC-MS m/z (ESI)= 267.10 [M+1]
Step 4: 7-(Ethyl-d5)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 one (105e)
2-(Methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 105d (2.5 g, 9.39 mmol) was dissolved in dimethylformamide (20 mL), and deuterated bromoethane (700 [L, 9.39 mmol) and sodium hydroxide (1.13 g, 28.16 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, then water was added and the organics were extracted with ethyl acetate and the organic layer was concentrated to obtain the title compound 7-(ethyl-d5)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl) 7,9-dihydro-8H-purin-8-one 105e (pale yellow solid, 1.65 g, 58.71% yield). LC-MS m/z(ESI)= 300.20 [M+1]
Step 7: 7-(Ethyl-d5)-2-(methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin 8-one (105f)
7-(Ethyl-d5)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 one 105e (1.6 g, 5.34 mmol) was dissolved in dichloromethane (8 mL), and m chloroperoxybenzoic acid (3.69 g, 21.38 mmol) was added with stirring at room temperature. The reaction mixture was reacted for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/ethyl acetate (v/v) = 1:1) to obtain the title compound 7-(ethyl-d5)-2-(methylsulfinyl)-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 105f (white solid, 1.4 g,
79.04% yield). LC-MS m/z(ESI)= 332.20 [M+1]
Step 8: 4-((7-(Ethyl-d5)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2 yl)amino)-2-fluoro-5-methylbenzamide (compound 105)
7-(Ethyl-d5)-2-(methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin 8-one 105f (300 mg, 0.91 mmol), 4-amino-3-methylbenzamide intermediate 2 (457 mg, 2.72 mmol) and potassium tert-butoxide (305 mg, 2.72 mmol) were dissolved in dimethylformamide (10 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 20:1) to obtain the title compound 4-((7-(ethyl-d5) 8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5 methylbenzamide compound 105 (white solid, 45 mg, 11.85% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.54 (s, 1H), 8.25 (s, 1H), 7.89 (d, 1H), 7.54 (d, 1H), 7.47 (s, 1H), 7.39 (s, 1H), 4.49-4.38 (m, 1H), 3.98 (dd, 2H), 3.47-3.38 (m, 2H), 2.59 2.51 (m, 2H), 2.29 (s, 3H), 1.73-1.65 (m, 2H). LC-MS m/z(ESI)= 420.20 [M+1]
Example 106 4-((7-(2,2-Difluoroethyl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)-2-fluoro-5-methylbenzamide (compound 106)
F 0 r
H2 N N N H
Compound 106 F F F H F F 0 N N -~ NN NF
N =- StepI Nep Step2tp 3 H2N N 0F N NN N H
0 0 0 0o
105d 106a 106b Compound 106 Step 1: 7-(2,2-Difluoroethyl)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H purin-8-one (106a)
2-(Methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 105d (1 g, 2.53 mmol) was dissolved in dimethylformamide (5 mL), and allyl iodide (397 [L, 4.51 mmol) and potassium carbonate (1.56 g, 11.26 mmol) were added with stirring at room temperature. The reaction mixture was stirred for 1 h, then heated to 65 °C and stirred for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, then water was added to the reaction mixture, and a solid was precipitated. The solid was collected by filtration and washed with water and petroleum ether, and concentrated by rotary evaporation under reduced pressure to obtain the title compound 7-(2,2 difluoroethyl)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8 one 106a (white solid, 800 mg, 64.49% yield). 1H NMR (600 MHz, DMSO-d6) 6 8.36 (s, 1H), 6.52-6.22 (m, 1H), 4.50-4.43 (m, 1H), 4.38-4.30 (m, 2H), 3.98 (dd, 2H), 3.49-3.42 (m, 2H), 2.53 (s, 3H), 2.50-2.45 (m, 2H), 1.79-1.60 (m, 2H). LC-MS m/z(ESI)= 331.10 [M+1]
Step 2: 7-(2,2-Difluoroethyl)-2-(methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro 8H-purin-8-one (106b)
7-(2,2-Difluoroethyl)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H purin-8-one 106a (800 mg, 2.42 mmol) was dissolved in dichloromethane (6 mL), and m-chloroperoxybenzoic acid (543 mg, 3.15 mmol) was added with stirring at room temperature. The reaction mixture was stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, saturated sodium bicarbonate and dichloromethane were added to the reaction mixture for extraction, and the organic layer was concentrated to obtain the title compound 7-(2,2-difluoroethyl)-2 (methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 106b (white solid, 740 mg, 88.23% yield). LC-MS m/z(ESI)= 347.10 [M+1]
Step 3: 4-((7-(2,2-Difluoroethyl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin 2-yl)amino)-2-fluoro-5-methylbenzamide
7-(2,2-Difluoroethyl)-2-(methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro 8H-purin-8-one 106b (200 mg, 0.58 mmol), 4-amino-3-methylbenzamide intermediate 2 (388 mg, 2.31 mmol) and cesium carbonate (564 mg, 1.73 mmol) were dissolved in dimethylformamide (5 mL). The reaction mixture was heated to 80 °C and stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with dichloromethane. The organic layer was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 30:1), followed by recrystallization to obtain the title compound 4-((7-(2,2-difluoroethyl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9 dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 108 (white solid, 65 mg, 24.99% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.64 (s, 1H), 8.21 (s, 1H), 7.84 (d, 1H), 7.54 (d, 1H), 7.47 (s, 1H), 7.41 (s, 1H), 6.54-6.22 (m, 1H), 4.51-4.41 (m, 1H), 4.37-4.25 (m, 2H), 3.98 (dd, 2H), 3.46-3.38 (m, 2H), 2.58-2.50 (m, 2H), 2.29 (s, 3H), 1.75-1.67 (m, 2H). LC-MS m/z(ESI)= 451.20 [M+1]
Example 107 (S)-4-((7-Ethyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 107)
0 1f H 2N N N
0 Compound 107 H 0
S- Step 1,I Step 2 H2N N N CIN NI N _________ N N
0 00 76c 107a Compound 107 Step 1: (S)-2-Chloro-7-ethyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (107a)
(S)-2-Chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 76c (2 g, 8.31 mmol) was dissolved in dimethylformamide (20 mL), and cesium carbonate (2.71 g, 8.31 mmol) and iodoethane (1.56 g, 9.97 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, then the reaction mixture was poured into 60 mL of water, and a large amounts of solid was precipitated The solid was collected by filtration and dried to give the title compound (S)-2-chloro-7-ethyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H purin-8-one 107a (white solid, 1.1 g, 49.26% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.43 (s, 1H), 5.04-4.97 (m, 1H), 4.15-4.06 (m, 1H), 3.98 (t, 1H), 3.91-3.84 (m, 4H), 2.44-2.37 (m, 1H), 2.30-2.19 (m, 1H), 1.26 (s, 3H). LC-MS m/z(ESI)= 269.10 [M+1]
Step 2: (S)-4-((7-Ethyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 107)
(S)-2-Chloro-7-ethyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 107a (500 mg, 1.86 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (1.25 g, 7.44 mmol), cesium carbonate (2.43 g, 7.44 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (253 mg, 0.28 mmol) were dissolved in 1,4-dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1) followed by Pre-HPLC to obtain the title compound (S)-2-fluoro-5-methyl-4-((7-methyl-8-oxo 9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 107 (white solid, 120 mg, 16.11%yield). 'H NMR (400 MHz, DMSO-d6) 6 8.56 (s, 1H), 8.27 (s, 1H), 7.88 (d, 1H), 7.54 (d, 1H), 7.47 (s, 1H), 7.41 (s, 1H), 5.01-4.94 (m, 1H), 4.09 (q, 1H), 3.96 (t, 3H), 3.92-3.82 (m, 4H), 2.76-2.66 (m, 1H), 2.33-2.23 (m, 1H), 2.28 (s, 3H), 1.24 (t, 3H). LC-MS m/z(ESI)= 401.20 [M+1]
Example 108 (R)-4-((7-Ethyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 108)
0
H2N : Or HF NNN N H ~ (R)
Compound 108 C H 0 N N~ N1 Step N , Co Step 2 H2N N NO CI S(R) CIN (R) - F NN N C C3 H
75c 108a Compound 108 0
Step 1: (R)-2-Chloro-7-ethyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one (108a)
(R)-2-Chloro-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 75c (1.4 g, 5.82 mmol) was dissolved in dimethylformamide (20 mL), and cesium carbonate (1.90 g, 5.82 mmol) and iodoethane (998 mg, 6.40 mmol) were added at 0 °C. The reaction mixture was stirred for 1 h. The reaction was monitored by TLC. After completion of the reaction, then the reaction mixture was poured into 60 mL of water, and a large amounts of solid was precipitated. The solid was collected by filtration and dried to yield the title compound (R)-2-chloro-7-ethyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro 8H-purin-8-one 108a (white solid, 1.08 g, 69.09% yield). LC-MS m/z(ESI)= 269.10 [M+1]
Step 2: (R)-4-((7-Ethyl-8-oxo-9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 108)
(R)-2-Chloro-7-ethyl-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8-one 108a (300 mg, 1.12 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (939 mg, 5.58 mmol), cesium carbonate (1.09 g, 3.35 mmol) and methanesulfonato(2 dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino 1,1'-biphenyl-2-yl)palladium(II) (202 mg, 0.22 mmol) were dissolved in 1,4-dioxane (10 mL), followed by nitrogen purging. The reaction mixture was stirred under nitrogen atmosphere at 110 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane/methanol (v/v) = 20/1) followed by Pre-HPLC to obtain the title compound (R)-2-fluoro-5-methyl-4-((7-methyl-8-oxo 9-(tetrahydrofuran-3-yl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 108 (white solid, 100 mg, 22.37% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.53 (s, 1H), 8.28 (s, 1H), 7.87 (d, 1H), 7.55 (d, 1H), 7.47 (s, 1H), 7.41 (s, 1H), 5.03-4.97 (m, 1H), 4.11-4.05 (m, 1H), 3.96 (t, 3H), 3.94-3.85 (m, 4H), 2.77-2.70 (m, 1H), 2.36-2.23 (m, 1H), 2.28 (s, 3H), 1.26 (t, 3H). LC-MS m/z(ESI)= 401.20 [M+1]
Example 109 2-Fluoro-4-((7-(fluoromethyl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 109)
F
0 4 H2 N N N
Fa N LN N H
Compound 109 F F F
H 0
Ss step 1 s Step 2 Step 3 H2 N N Sn" N)o S'I:W:N=0 F : N NN H
0 00
105d 109a 109b Compound 109 Step 1: 7-(2-Fluoroethyl)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin 8-one (109a)
2-(Methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 105d (1.0 g, 3.75 mmol), 1-fluoro-2-iodoethane (0.71 g, 4.1 mmol) and potassium carbonate (1.5 g, 11.2 mmol) were added to a dry reaction flask, followed by the addition of DMF (10 mL). The reaction mixture was stirred at room temperature for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with dichloromethane, and the organic phases were combined, dried and concentrated to yield the title compound 7-(2-fluoroethyl)-2-(methylthio)-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 109a (white solid, 1.0 g, 85.47% yield). LC-MS m/z (ESI) = 313.10 [M+1]
Step 2: 7-(2-Fluoroethyl)-2-(methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H purin-8-one (109b)
7-(2-Fluoroethyl)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin 8-one 109a (0.9 g, 2.88 mmol) was added to a dry reaction flask and dissolved in DCM (10 mL), and m-chloroperoxybenzoic acid (0.5 g, 3.17 mmol) was added in three batches at room temperature. After addition, the reaction mixture was stirred for 3 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with dichloromethane, and the organic phases were combined, dried and concentrated to yield the title compound 7-(2-fluoroethyl)-2 (methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 109b (white solid, 0.8 g, 85.11% yield). LC-MS m/z (ESI) = 329.10 [M+1]
Step 3: 2-Fluoro-4-((7-(fluoromethyl)-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H purin-2-yl)amino)-5-methylbenzamide (compound 109)
7-(2-Fluoroethyl)-2-(methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H purin-8-one 109b (200 mg, 0.61 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (410 mg, 2.44 mmol) and cesium carbonate (590 mg, 1.83 mmol) were added to a dry reaction tube, followed by the addition of DMF (3 mL). The reaction mixture was stirred at 80 °C for 2.5 h. Water was added and the organics were extracted with dichloromethane, and the organic phases were combined, dried and concentrated. The residue was purified by silica gel column chromatography (DCM/MeOH (v/v) = :1) to obtain the title compound 2-fluoro-4-((7-(fluoromethyl)-8-oxo-9-(tetrahydro 2H-pyran-4-yl)-8,9-dihydro-7H-purin-2-yl)amino)-5-methylbenzamide compound 109 (white solid, 21 mg, 7.98% yield). 'H NMR (600 MHz, DMSO-d6) 6 8.60 (s, 1H), 8.21 (s, 1H), 7.87 (d, 1H), 7.55 (d, 1H), 7.48 (s, 1H), 7.42 (s, 1H), 4.75-4.66 (m, 2H), 4.48-4.43 (m, 1H), 4.19-4.13 (m, 2H), 4.00-3.97 (m, 2H), 3.43 (t, 2H), 2.59-2.53 (m, 2H), 2.29 (s, 3H), 1.74-1.68 (m, 2H). LC-MS m/z (ESI) = 433.20 [M+1]
Example 110 2-Fluoro-5-methyl-4-((8-oxo-9-(tetrahydro-2H-pyran-4-yl)-7-(2,2,2-trifluoroethyl) 8,9-dihydro-7H-purin-2-yl)amino)benzamide (compound 110)
F
o F F
H2N N NO
H
0
Compound 110
F F F F F H F S O N N Step 2 Step 3 H 2N NN N
a H
105d 110a 110b Compound 110 Step 1: 2-(Methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7-(2,2,2-trifluoroethyl)-7,9-dihydro-8H purin-8-one (110a)
2-(Methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 105d (1.0 g, 3.75 mmol), 2-iodo-1,1,1-trifluoroethane (444 mg, 4.5 mmol) and cesium carbonate (3.67 g, 11.26 mmol) were added to a dry reaction flask, followed by the addition of DMF (10 mL). The reaction mixture was stirred at room temperature for 2.5 h. The reaction was monitored by TLC. After completion of the reaction, water was added and the organics were extracted with ethyl acetate, and the organic phases were combined, dried and concentrated to give the title compound 7-(2-fluoroethyl)-2-(methylthio)-9 (tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one 110a (pale yellow solid, 1.18 g, 90.22% yield). H NMR (600 MHz, DMSO-d6) 6 8.43 (s, 1H), 4.83 (q, 2H), 4.52-4.43 (m, 1H), 3.98 (dd, 2H), 3.51-3.42 (m, 2H), 2.53 (s, 3H), 2.50-2.44 (m, 2H), 1.76-1.68 (m, 2H). LC-MS m/z (ESI) = 349.10 [M+1]
Step 2: 2-(Methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7-(2,2,2-trifluoroethyl)-7,9-dihydro 8H-purin-8-one (11Ob)
7-(2-Fluoroethyl)-2-(methylthio)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin 8-one 110a (1.17 g, 3.36 mmol) was added to a dry reaction flask and dissolved in DCM (10 mL), and m-chloroperoxybenzoic acid (754 mg, 4.37 mmol) was added at room temperature. The reaction mixture was stirred for 1.5 h. The reaction was monitored by TLC. After completion of the reaction, saturated sodium bicarbonate and dichloromethane were added to the reaction mixture for extraction, and the organic phases were combined, dried and concentrated to obtain the title compound 2 (methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7-(2,2,2-trifluoroethyl)-7,9-dihydro 8H-purin-8-one 11Ob (white solid, 1 g, 81.72% yield). LC-MS m/z (ESI) = 365.10 [M+1]
Step 3: 2-Fluoro-5-methyl-4-((8-oxo-9-(tetrahydro-2H-pyran-4-yl)-7-(2,2,2-trifluoroethyl) 8,9-dihydro-7H-purin-2-yl)amino)benzamide (compound 110)
2-(Methylsulfinyl)-9-(tetrahydro-2H-pyran-4-yl)-7-(2,2,2-trifluoroethyl)-7,9-dihydro 8H-purin-8-one 110b (200 mg, 0.55 mmol), 4-amino-2-fluoro-5-methylbenzamide intermediate 2 (369 mg, 2.20 mmol) and cesium carbonate (537 mg, 1.65 mmol) were added to a dry reaction tube, followed by the addition of DMF (3 mL). The reaction mixture was stirred at 40 °C for 5 h. Water was added and the organics were extracted with dichloromethane, and the organic phases were combined, dried and concentrated. The residue was purified by silica gel column chromatography (DCM/MeOH (v/v) = :1) to obtain the title compound 2-fluoro-5-methyl-4-((8-oxo-9-(tetrahydro-2H pyran-4-yl)-7-(2,2,2-trifluoroethyl)-8,9-dihydro-7H-purin-2-yl)amino)benzamide compound 110 (white solid, 45 mg, 17.5% yield). 'H NMR (600 MHz, DMSO-d6) 6 8.54 (s, 1H), 8.26 (s, 1H), 7.87 (d, 1H), 7.55 (d, 1H), 7.47 (s, 1H), 7.42 (s, 1H), 4.85-4.69 (m, 2H), 4.53-4.42 (m, 1H), 3.98 (dd, 2H), 3.46 3.40 (m, 2H), 2.52-2.45 (m, 2H), 2.29 (s, 3H), 1.76-1.68 (m, 2H). LC-MS m/z (ESI) = 469.20 [M+1]
Example 111 4-((9-(cis-3-Cyanocyclobutyl)-7-ethyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 111)
0 H2 N N NO Fa N<N N
CN
Compound 111
H f N N NN~ S o step 1 O Step 2 O Step 3 H2 N N N SN SS N N F N n~' N>
CO CN CN H CN 74e 111a 111b Compound 111
Step 1: cis-3-(7-Ethyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (111a)
cis-3-(2-(Methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 74e (500 mg, 19.1 mmol) was dissolved in dimethylformamide (5 mL), and iodoethane (3.0 g, 19.1 mmol) and sodium hydroxide (1.53 g, 38.2 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, Then 50 mL of water was added, and a solid was precipitated. The solid was collected by filtration and dried by rotary evaporation under reduced pressure to obtain the title compound cis-3-(7-ethyl-2-(methylthio)-8-oxo-7,8 dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 111a (white solid, 487 mg, 87.9% yield). 1H NMR (400 MHz, DMSO-d6) 6 8.68 (s, 1H), 4.97-4.87 (m, 1H), 3.93 (q, 2H), 3.34 (s, 3H), 3.33-3.23 (m, 2H), 2.91 (s, 3H), 2.77-2.69 (m, 2H), 1.27 (t, 3H). LC-MS m/z (ESI)= 290.10 [M+1]
Step 2: cis-3-(7-Ethyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile (111b)
cis-3-(7-Ethyl-2-(methylthio)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 111a (487 mg, 1.66 mmol) was dissolved in methanol (10 mL), and 2 mL of purified water was added, followed by the addition of potassium peroxymonosulfate (1.02 g, 1.66 mmol). The reaction mixture was heated to 60 °C and stirred for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated by rotary evaporation under reduced pressure to remove the methanol, and then 5 mL of purified water was added. The mixture was extracted 3 times with dichloromethane (20 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate and concentrated by rotary evaporation under reduced pressure to remove the solvent and thus to obtain cis-3-(7-ethyl-2-(methylsulfonyl)-8 oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1-carbonitrile 111b (pale yellow solid, 397 mg, 74.3% yield). LC-MS m/z(ESI)= 322.10 [M+1]
Step 3: 4-((9-(cis-3-Cyanocyclobutyl)-7-ethyl-8-oxo-8,9-dihydro-7H-purin-2-yl)amino)-2 fluoro-5-methylbenzamide (compound 111)
cis-3-(7-Ethyl-2-(methylsulfonyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclobutane-1 carbonitrile 111b (200 mg, 0.62 mmol) and 4-amino-3-methyl-2-fluorobenzamide intermediate 2 (538 mg, 3.20 mmol) were dissolved in 3 mL of N,N dimethylformamide, followed by nitrogen purging and the addition of potassium tert butoxide (143 mg, 1.28 mmol). The reaction mixture was stirred under nitrogen atmosphere at room temperature for 4 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into 30 mL of purified water, and a large amounts of yellow solid was precipitated. The solid was collected by filtration and purified by silica gel column chromatography (dichloromethane/methanol (v/v)= 35/1) to obtain the title compound 4-((9-(cis-3-cyanocyclobutyl)-7-ethyl-8-oxo 8,9-dihydro-7H-purin-2-yl)amino)-2-fluoro-5-methylbenzamide compound 111 (white solid, 25 mg, 10.0% yield). 'H NMR (400 MHz, DMSO-d6) 6 8.63 (s, 1H), 8.28 (s, 1H), 7.88 (d 1H), 7.58-7.56 (m, 1H), 7.50 (s, 1H), 7.41 (s, 1H), 5.14-5.09 (m, 1H), 3.87-3.82 (m, 2H), 3.64-3.59 (m, 1H), 3.26-3.16 (m, 2H), 2.75-2.68 (m, 2H), 2.32 (s, 3H), 1.22 (t, 3H). LC-MS m/z(ESI)= 410.20 [M+1]
Biological experiments
1. Inhibition assay of DNA-PK kinase
The inhibitory activity of the compounds on DNA-PK kinase was measured by using a DNA-PK kinase assay kit (purchased from Promega, Cat.#: V4107, Lot: 0000366495). The results were quantified by chemiluminescence, and the detailed experimental protocol was as follows: i. An ADP-fluorescence standard curve over a range of concentrations was plotted according to the kit instructions; ii. 5 L reaction systems were prepared in a 384-well white plate, and to each well 1 L of a compound (each under a concentration gradient over 1 M, 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM, 0.064 nM, 0.013 nM), 20 units of DNA-PK kinase, 0.2 [g/[tL substrate, 10 [g/[tLDNA, 50 [MATP, and 1% DMSO were added; iii. The mixtures were mixed well, centrifuged (1000 rpm, 30 s), and incubated at 37°C for 60 min; iv. 5 pL of ADP-GloTMReagent was added to stop the reaction, which was then mixed well, centrifuged (1000 rpm, 30 s), and incubated at room temperature for 40 min; v. 10 pL Kinase Detection Reagent was added, and the mixture was mixed well by shaking, centrifuged (1000 rpm, 30 s), and incubated at room temperature for 30 min; vi. The fluorescence value was measured by using a microplate reader (Thermo fisher, Varioskan LUX). IC 5 o was calculated using GraphPad Prism 8, and the results are shown in Table 1.
Table 1. Inhibitory activity of compounds of the present disclosure on DNA-PK kinase Compound No. IC 5 0(nM) 1 10.10 2 0.08 3 18.58 4 8.30 5 0.78 6 0.78
7 5.47 8 11.70 9 0.47 11 25.03 5.28 17 28.66 18 2.10 21 53.00 22 1.36 24 0.56 3.44 26 7.60 27 4.36 29 3.90 66.60 31 31.00 36 17.95 37 11.88 38 0.47 39 5.70 39.2 41 13.42 42 2.05 43 9.80 44 1.10 13.53 46 0.99 47 3.69 48 1.96 12.72 51 34.09 53 0.32 54 8.07 14.76 56 14.33 57 3.98 59 12.12 29.74 62 4.17 63 50.47 64 3.34 67 2.96 68 0.01 69 1.25 4.14 71 2.17 72 0.05 73 1.34 5.69 76 1.96 77 4.67 78 0.038 79 1.86 11.15 81 47.12
82 24.5 83 0.43 84 0.24 85 1.10 86 0.44 88 2.85 89 3.83 90 10.82 91 2.44 92 0.02 93 0.75 94 7.20 95 0.60 96 3.89 97 0.68 98 4.01 99 2.13 100 2.20 101 9.60 105 5.54 106 6.75 107 10.11 109 5.54 103 (Comparative Example) 100.40
The results show that the compounds of the present disclosure have a good inhibitory effect on DNA-PK kinase.
2. Metabolic stability in liver microsomes
1.1 Experimental materials Liver microsomes (purchased from Reed Liver Disease Research (Shanghai) Co., Ltd., product: human liver microsomes (NJT)); NADPH (purchased from Roche, Lot: 40939); UDPGA; the compounds of the present invention (test substances).
1.2 Experimental procedure (1) A mixed working solution of testosterone (0.5 M) and 7-hydroxycoumarin (0.5 pM) was prepared. (2) A mixed solution of NADPH (1 mM) and UDPGA (1 mM) was prepared, and pre warmed at 37°C in a water bath shaker before use. (3) Liver microsomes were thawed and diluted with a buffer. (4) A preparation of a test substance (0.5 M) and a solution of liver microsomes (0.5 mg/mL) were added to each well of an incubation plate. (5) Three experimental groups were set and samples were added. Test group: the test substance was incubated with liver microsomes for 0 min, 30 min, 60 min, and 120 min in the presence of the coenzyme NADPH and UDPGA; Positive control group: testosterone (0.5 [M) and 7-hydroxycoumarin (0.5 [M) were incubated with microsomes for 0 min and 120 min in the presence of the coenzyme NADPH and UDPGA; Negative control group: the test substance was incubated with microsomes for 0 and 120 min without any coenzyme. (6) All samples were incubated at 37°C, and pre-cooled methanol was added at each termination time (0, 15, 30, 45, 60 min, 120 min) to terminate the reaction, and the time was recorded. (7) All samples were mixed well and centrifuged at 4000 rpm for 10 min. The supernatant was taken and measured by LC-MS/MS for the amount of the test substance remaining in the samples. Data analysis was carried out as follows: T 1/2=0.693/k, Clint (0.693/Ti1/2 ) x (1/ (microsomal protein ) x Scaling Factors), =
Note: k is the slope of the logarithm of the remaining amounts of compound versus time. The results are shown in Table 2.
Table 2. Stability of compounds of the present disclosure in human liver microsomes Compounds T/2 (min) Compound 2 >120 Compound 5 >120 Compound 8 >120 Compound 9 >120 Compound 18 >120 Compound 24 >120 Compound 25 >120 Compound 26 >120 Compound 27 >120 Compound 29 >120 Compound 38 >120 Compound 39 >120 Compound 42 >120 Compound 44 >120 Compound 48 >120 Compound 53 >120 Compound 54 >120 Compound 57 >120 Compound 69 >120 Compound 72 >120 Compound 73 >120 Compound 75 >120 Compound 76 >120 Compound 77 >120 Compound 79 >120 Compound 80 >120 Compound 81 >120 Compound 82 >120 Compound 83 >120 Compound 85 >120 Compound 92-1 >120 Compound 92-2 >120 Compound 93 >120
The results show that the compounds of the present application have good stability in human liver microsomes.
3. Pharmacokinetic assay
3.1 Experimental materials ICR mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.
3.2 Experimental procedure (1) Healthy male ICR mice (18-22 g) were prepared, with 18 mice for each compound. The mice were divided into 2 groups (an i.v. group and a p.o. group), with 9 mice per group. 3 mice were picked for blood collection at each time point. (2) After the animals were fasted overnight (with free access to water), the compound of the present invention dissolved (or suspended) in a solvent of 5% DMSO and 95% % HP-P-CD (v:v) was administered via the tail vein (i.v., 1 mg/kg) or intragastrically (p.o., 10 mg/kg), respectively. (3) For the i.v. group, 0.1 mL blood was collected from the submandibular vein 5 min, min, 0.5 h, 1 h, 2 h, 4 h, 8 h,12 h, and 24 h after administration, respectively, anticoagulated with EDTA-K2, and centrifuged at 4°C for 5 min to obtain plasma, which was stored at -20°C until testing. (4) For the p.o. group, 0.1 mL blood was collected from the submandibular vein before administration and 15 min, 0.5 h, h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h after 1 administration, and treated in the same manner as that for the intravenous group. (5) The concentration of the compound of the present invention in plasma was measured by LC/MS/MS. (6) Calculation and fitting were performed with Analyst 1.6 from AB Company, and the results are shown in Table 3.
Table 3. Pharmacokinetic results of the compounds of the present disclosure
Compounds Administration AUC Cmax TI(h) F% Route (h*ng/mL) (ng/mL) Compound 2 iv 265 44 38.2 po1044 449 1.84 3. Compound 5 iv 234 - 0.333 po2266 835 3.21 Copud6iv 82.4 - 0.299 17 Compound6o 1482 1220 0.964 Copud9iv 323 - 0.41 5. Compound 1659 1487 1.3451.4 Compound 18 iv 171 - 0.771 78.4 po1362 312 3.06 Compound 24 iv 466 0.95 135 _19 6276 2500 - 3.73
Cmon26iv 639 -1.5210 Compound26 7104 2733 2.9 110 Cmon27iv 679 - 1.48 21 Compound27o 14582 5450 2.17213 Compound 29 iv 609 - 0.46 - 42.1 Co 2562 1820 1.22 Compound 38 iv 269 - 0.341 - 4. Co 1308 2203 1.02 Compound 41 iv 357 - 0.55 42.3 po1505 2713 4.88 Cmon42iv 230 - 1.41 - 7 Compound42o 1499 858 1.7370 Compound 54 iv 840 - 5.22 - 63.9 Co 5369 2057 3.21 Compound 62 iv 1191 - 1.69 44.8 po4717 1853 3.01 Cmon72iv 1698 - 2.92 16 Compound72o 28250 4680 2.58166 Compound 73 iv 239 - 1.24 56 po1274 484 2.50 Cmon75iv 109 - 0.31 - 6. Compound75 756 984 2.1069.2 Compound 76 iv 120 - 0.30 - 104 Co 1365 1571 0.93 Compound 77 iv 545 - 1.55 108 po5971 1747 3.2410 Compound 78 iv 216 - 1.17 65.2 po1204 446 3.5 Compound 79 iv43 2487 2.15 100
Cmon80iv 6299 - 3.12 - 3. Compound80o 21207 3570 2.1633.6
The results show that the compounds of the present disclosure have good pharmacokinetic properties.
4. TTK kinase activity assay
4.1 Experimental materials TTK kinase was purchased from Invitrogen, Cat.#: PV3792.
4.2 Experimental procedure (1) 1xbuffer was prepared by diluting 5xbuffer into 1xbuffer with ddH2 0, and adding 0.05 mM DTT. (2) The compounds of the present invention were prepared at an initial concentration of 1000 nM, and diluted by 1:3 to obtain 10 serial dilutions. (3) The diluted compounds of the present invention were added to a 384-well plate (purchased from Greiner, product number 784075) by an Echo 550 pipetting workstation, and centrifuged at 1000 g for 1 min.
(3) A 2xTTK enzyme solution was prepared with the 1xbuffer. Then, 2.5 L of the 2x enzyme solution was added to each well, centrifuged at 1000 g for 30s, and left at room temperature for 10 min. (4) A 2xMBP substrate/ATP mixture was prepared with the 1xbuffer. Then, 2.5 L of the 2xMBP substrate/ATP mixture was added to each well, centrifuged at 1000 g for s, and left at room temperature for 1h. (5) 5 pL ADP-Glo was added to each well and left at room temperature for 40 min. (6) 10 L of the detection reagent was added to each well and left at room temperature for 40 min. (7) The results were read under an Envision 2104 microplate reader, and curve fitting and IC 5 o calculation were performed with GraphPad Prism 8. The results are shown in Table 4.
Table 4. TTK kinase inhibiting activity of compounds of the present disclosure Compound No. IC5 0 (nM) Compound 2 275.2 Compound 5 1722 Compound 9 10000 Compound 18 386.6 Compound 24 4219 Compound 27 427.9
The results show that the compounds of the present disclosure have no significant inhibitory effect on TTK kinase, but have high selectivity for DNA-PK kinase.
Some specific embodiments have been described in detail in the specification of the present invention. Those skilled in the art should realize that these embodiments are exemplary and not to be construed as limiting the present invention. For those skilled in the art, various improvements and modifications may be made to the present invention without departing from the principles of the present invention, and the technical solutions obtained by such improvements and modifications also fall within the scope of the claims of the present invention.

Claims (18)

Claims
1. A compound of general formula (I), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, RO R3 NN O
(R C NH N
(I) wherein R3R3R B R3I
or(R2 R (I is ( 3R 2
--- is a single bond or a double bond; A R3 B, R2 nC A, B, C, and D in the are each independently C or N, and at least one of A, B, C, and D is N; Ro is H, C1 -6 alkyl, or cyclopropyl, wherein the C1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium (D);
0 Ri is 0, , 0, ,0, Rib
RO N R~, Rlb Rlb ,0 X , N Rla or pyridyl, and Ri is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, C1-6 alkyl, and C1-6 alkoxy;
Ria is H or C 1 -6 alkyl; Rib is H, OH, cyano, or hydroxyl-substituted CI-6 alkyl; R 2 is H, cyano, =0, carboxyl, -C(=)NR2aR2b, C1-6 alkoxy, C1-6 alkyl, halogen, S(=0) 2R2 a or a -C(=O)OC 1 -6 alkyl; wherein the CI6 alkyl, -C(=O)OC 1 .6 alkyl or Ci-6 alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium;
R2 a and R2b are each independently H, C1-6 alkyl or 3- to 5 -membered cycloalkyl, wherein the C1 .6 alkyl is optionally further substituted with one or more substituents selected from OH, D, halogen, C1 .6 alkyl and C1 .6 alkoxy; alternatively, R 2 a and R2b together with the atom to which they are attached form a 5- to 6-membered heterocyclyl group containing 1, 2 or 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from C1-6 alkyl, OH, and halogen; R3 is halogen or C1 .6 alkyl, wherein the C1-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; and x and y are each independently 1, 2, or 3; provided that R3
RH C R 1 is not , , or OH when , Ro, R2 , and R3 all satisfy the following condition: R3
( 2 (R is ,n is 1, Ro is H or methyl, R2 is methoxy or S(=0)2Me, and R3 is methyl.
2. The compound according to claim 1, or a stereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt, or co-crystal thereof, wherein
F OH
Ri is OH , OH , OH F
D OH CN, OH CN CNO
0 0,O ~ 0ll Ria OH 0~"
0 OH OH OH , OH
7 0 , CN,or O O, N Ria is H or C1 -6 alkyl; R2 is H, cyano, -C(=0)NR2aR2b, C1-6 alkoxy, halogen, -S(=0) 2 R2 a or -C(=O)OCi 6alkyl; wherein the -C(=O)OCI-6 alkyl or C1 -6alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2 a and R2b are each independently H, C 1-6 alkyl or 3- to 5-membered cycloalkyl, wherein the C1 -6 alkyl is optionally further substituted with one or more substituents selected from OH, D, and halogen; alternatively, R 2 a and R2b together with the atom to which they are attached form a 5- to 6-membered heterocyclyl group containing 1 to 3 heteroatoms selected from N, o and S, wherein the heterocyclyl group is optionally further substituted with one or more substituents selected from OH and halogen; R3 is halogen or C1 -6alkyl, wherein the CI-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; mis0or 1; and n is 0, 1, or 2; provided that R3
(R2 C R 1 is not , , or OH when , Ro, R2 , and R3 all satisfy the following condition: R3
( 2 (R is , n is 1, Ro is H or methyl, R2 is methoxy or S(=0)2Me, and R 3 is methyl.
3. The compound according to claim 1, or a stereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt, or co-crystal thereof, wherein R3
(Ri (R 2 3 or (R 2
Ro is H, CiA alkyl, or cyclopropyl, wherein the Ci4 alkyl is optionally further substituted with one or more substituents selected from halogen and D;
Riis O, 0 ,0 Ria ,OH, 0 o DO CN , CN, CN , O-~, OH , O, or O0 Ria is H, a C1-6 alkyl or a -C(=)C1-6 alkyl; R 2 is H, cyano, -C(=0)NR2aR2b, a C1-6 alkoxy, halogen, -S(=0)2R2a or a C(=O)OCI-6 alkyl; wherein the -C(=O)O alkyl or CI-6 alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2 a and R2b are each independently H, CI-6 alkyl or 3- to 5-membered cycloalkyl, wherein the C1 -6 alkyl is optionally further substituted with one or more substituents selected from OH, D, and halogen; alternatively, R 2 a and R2b together with the atom to which they are attached form - to 6-membered heterocyclyl group containing 1 to 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from OH and halogen; R3 is halogen or C1 -6 alkyl, wherein the CI-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; mis0or 1; and n is 0, 1, or 2; provided that R3
R C R 1 is not , , or OH when , Ro, R2 , and R3 all satisfy the following condition: R3
(R 2 C (R2 R3 is , n is 1, Ro is H or methyl, R2 is methoxy or S(=0)2Me, and R3 is methyl.
4. The compound according to claim 1, or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, wherein the compound is a compound of general formula (II):
RO R2d ~ R3 N R2C N N N H R1 (20) wherein Ro is H, C1 -6 alkyl, or cyclopropyl, wherein the C1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
0O Ri is 0, , , 0, 0, R1b
O N R1 b, R1 b, R1b ,, X , Rla, or pyridyl, and Ri is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxyC-6alkyl, andC- 6 alkoxy; Ria is H orC1 -6alkyl; Ribis H, OH, cyano, or hydroxyl-substitutedC1-6alkyl; R2e is H, cyano, halogen, orC1-6alkoxy; R2d is H, cyano, carboxyl, -C(=)NR2aR2b, C1-6 alkyl, halogen, -S(=0)2 R 2 a or C(=O)OC 1-6alkyl; wherein theCI-6alkyl or-C(=0)OCi-6alkyl is optionally substituted with one or more substituents selected from halogen and deuterium; R2 a and R2bare each independently H,C 1-6alkyl or 3- to 5-membered cycloalkyl, wherein theCI-6 alkyl is optionally further substituted with one or more substituents selected from OH, D, halogen,Ci-6 alkyl andC1 -6alkoxy; alternatively, R 2 aand R2btogether with the atom to which they are attached form - to 6-membered heterocyclyl group containing 1 to 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected fromCi-6alkyl, OH, and halogen; R3 is halogen orCI-6 alkyl, wherein theCI 6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; and x and y are each independently 1, 2, or 3; provided that R3
RH C R 1 is not , , or OH when , Ro, R2 , and R3 all satisfy the following condition:
R3
(R C (R 2 R is selected from , n is 1, Ro is H or methyl, R2 is methoxy or -S(=0)2Me, and R 3 is methyl.
5. The compound according to claim 4, or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, wherein the compound is a compound of general formula (III): 0 RO R2 a\ 3 . N R2 b-N N R3 NRN R2C N N N H R1 (III) wherein Ro is H, C1 -6 alkyl, or cyclopropyl, wherein the C1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
0 s~ Ri is 0, , 0,,ORib
Oy N Rib Rib, Rib ( 0 X N Ra, or pyridyl, and Ri is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, C1-6 alkyl, and C1-6 alkoxy; Rib is H, OH, cyano, or hydroxyl-substituted C1-6 alkyl; R2 a and R2b are each independently H, CI-6 alkyl or 3- to 5-membered cycloalkyl, wherein the C1 -6 alkyl is optionally further substituted with one or more substituents selected from D and halogen; alternatively, R 2 aand R2b together with the atom to which they are attached form - to 6-membered heterocyclyl group containing 1 to 3 heteroatoms selected from N, 0 and S, wherein the 5- to 6-membered heterocyclyl group is optionally further substituted with one or more substituents selected from C1-6 alkyl, OH, and halogen; R2e is H, cyano, halogen, or CI-6 alkoxy, wherein the CI6 alkoxy is optionally substituted with one or more D; R3 is halogen or C1 -6 alkyl, wherein the C1-6 alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is 0 or 1; n is 0, 1, or 2; and x and y are each independently 1, 2, or 3.
6. A compound of general formula (IV), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, 0 RO H 2N N N F N N N H R1 (IV) wherein Ro is H, C 1-6 alkyl, or cyclopropyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium; and Ri is -(CH)m- 4 to 7-membered carbocyclyl, -(CH)m- 4 to 7-membered heterocyclyl, -(CH)m- 8 to 12-membered bridged ring, or -(CH)m- 7 to 12 membered spiro, wherein the -(CH)m- 4 to 7-membered carbocyclyl, -(CH)m- 4 to 7-membered heterocyclyl, -(CH)m- 8 to 12-membered bridged ring, or -(CH)m- 7 to 12 membered spiro is optionally further substituted with one or more substituents selected from hydroxy, cyano, halogen, =0, C1-6 alkyl, C 1-6 alkoxy, and hydroxyl-substituted C 1-6 alkyl.
7. The compound according to claim 6, or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, wherein Ro is H, C 1-6 alkyl, or cyclopropyl, wherein the C 1-6 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
Ri is 0, , , 0, 0, Rib
O N Rlb , Rlb , Rlb '.R1a, or pyridyl, and Ri is optionally further substituted with one or two substituents selected from D, halogen, cyano, hydroxy, C1-6 alkyl, and C1-6 alkoxy; Ria is H or C1-6 alkyl; Rib is H, OH, cyano, or hydroxyl-substituted C1-6 alkyl; mis0or 1; and x and y are each independently 1, 2, or 3.
8. The compound according to claim 7, or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, wherein Ro is C 1 -4 alkyl, wherein the C 1 -4 alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium; and
Ri is OqN OH O'OH0, 0
10CN, CN F OH OH F
0, Li >t, N OH 0
> OH OCN 00 OH 0
OH OH ,or N
9. The compound according to any one of claims Ito 8,or astereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt, or co-crystal thereof, wherein the compound is: F Co N/\_ 0 N 3 N \I 0 N. N NN N N N N N N N N~~ N N H bH bH bH b 0 , 0 ,0 / 0 / N./0
HN NN2N N,',_N= NN N N 0k N NN N >0= N A' N >=QH 2 N FN H bH bH bH b 0 o 0 0 0 /N~ 0/ 0 >4 N N>= N_0 N '~ NN - C>=O H N-Cn> SNN NF N N NN N N H bH bF H b 0 0 0
"/ 11Ni /_N>== N N^ K NN :N\H NN N- -N>==H 2 2N N N> N NNN F H bH bH bH b 0 0 0 0
0 NNN ' / 0/ H N NN N0 Cf N> H 2N NNN NN N ~A NNN N H bH Hb
0 0 0
N0 0
N N --NNN >z I NA L- >ss H NI z N N N N N N N N N H H bHb 0 0 0
0 0 NN /0 NN /N ~ CD 3 N N N N N HAL H IAN N"Q N N N NIN NN N H bH bHb 0 0 0 0N-. CD /N ~ H N ~ N N>= H2 ~N HN- NN- AN"N N Z '' N' N H N >=oN H N N
00 0 N-/ N.. N N NNN N NAL N= _ A' " N N N N' N/' N N"N\oN >= H _0H _0 4z N N N ' N H N N
H H S,
N OH, 0, 0 0
N FN N N_ CFN /i N - \_ \/ 2N H N NNN " N NNN N NNNF NNN
H bH bNA Hb
0, 0, 0, N
0 0/ / H2N N H2N) -' N N F N N N = >=o AAN > F AN N H 2N N N >== H F N -N N
0 0 N 00/ 0 N N _D N N N N / NNF D 3C.' N N N N N = ~ N N 3C' '
H H H HH
0 0 0 0
0O/ 0 /H H 2N N1- NN AN N N N N7 ,N>= 0 N l'Z N>= A' N N/ N I >=" K-A- I NN N:NA' N N N N N N H bH bH bHb 0 0 0 0
N ININ IN N N N - SN
N NAN N N N N"N H bH H 0 0 0
HO S NNNH~~ NN 0 N
N N bH ~n N > N N IN H b 1N Hb
0 0 0 I 0 0/ S : >= NN NN N NN H 2N N >=N HN- AL N>= N N N N N N N N N H boH bHb O0 0
CD 3 /0/0/ H2N ~N IN D3CN N N N N0 N N IN H NI Nzz IN A> IF IN N NN~ N N N N IN H H b N Hb bO 0 0 / 0 / 0 /0 N IN N HO N IN N N N 112 IN IN
- '" NN\-0N N F CN N INN H bN~ N N N H b 3 Hb
0 0 0 0 0 /0 /0/ H 2N N, IN 0 F - N' N/ HOF NN
HO N N IN IN AN ~N F N N FN N ~N IN H boH bHb O0 0 0 0 // 0 11 2N - IN N IN -N N1 \ 0 N- N N>7:, 0 2 N - N N A >_\_ >-N N-; N H2 N~ >-o>0 N N~ H bH bH bHb 0
0 0 0 0 0 /0 00 H2N' - IN IN H 2N NN > H 2N -~ N N N AN N N>-o FNNN F NN NFN NN H ;jH bHb bo0 0 0 0 /0/
H12N-J N-1 N>= H1 2N4- N NNN _'_ F I = 2N N N N INN A- >zO H H F IN IN IN /H O 0 N-b
0/ 0 /0 /H 2 N N N \_N H2N' N N/ 2 N
NH F N N NH HH
tOHl 0
0 0 0
H.' F H NF HN N N> F N N, N
N N OH H 0/ 0 /H 2Nt N N0 N2 NN HN NI N F N'N N>= H 2, N)X I C>= F NI N N
FOH H
0 0 H2N N N H2Nt N N -N F N N N 2 N 0 F N N N H N N H F
O OH OH 0 /0 0 H2N N 2N N N I~N NO 2NN _-0 F N N N F N N NJ. H N H
0 CN OH
00/ H0 N H2Nt N NN N/ H 2N N- N \ N"N ,:_: F2t N N N H F N NN OHH
F2 N N N H H 0
2 NN NNN' N H 20 0 NN H N 2
F N N Nj F f
H ~Hb 0OH 0
0 0o- 0F HH2N'Nr>F N F N I-N, NF N N
H2 N NH 2 N 0 H2 0K- >=o NNN F N N NF N NHNN NNN~ H F-H HH b N N H2'OH
209:>=
F 0 0 F D3CN. H2N "==- HNNN - N"- N>= F N N N N N H b H 0 , or 0
10. A pharmaceutical composition, comprising: (1) a compound according to any one of claims 1 to 9, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, (2) optionally one or more additional active ingredients; and (3) a pharmaceutically acceptable carrier and/or excipient.
11. Use of the compound according to any one of claims 1 to 9, or a stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the pharmaceutical composition according to claim 10, in the manufacture of a medicament for treatment of cancer; wherein the medicament for treatment of cancer is preferably a DNA-PK inhibitor.
12. A compound of general formula (I'), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, RO R3 NN O
(R C NH N R (I') wherein C is selected from phenyl and 6-membered monocyclic heterocyclic group, wherein the heterocyclic group may contain 1 to 3 heteroatoms selected from N; Ro is selected from H, C 1 .6 alkyl, and cyclopropyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
Ri is selected from 0, 0 o , , Ra,
D OH CN, CN , CN , OH
sO O, and ; Ria is selected from H, C1 .6 alkyl, and -C(=O)C 1 .6 alkyl;
R2 is selected from cyano, carboxyl, -NR2aR2b, -C(=0)NR2aR2b, CI- 6 alkoxy, C 1-6 alkyl, halogen, -S(=0) 2 R2 aand -C(=)OC1-6 alkyl; wherein the alkyl or alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C1 -6 alkyl and 3- to 5-membered cycloalkyl, wherein the C1 -6 alkyl is optionally further substituted with one or more substituents selected from OH, D, halogen, C1 -6 alkyl and C1 -6 alkoxy; R2 a and R2b together with the atom to which they are attached may form 5- to 6 membered heterocyclyl group which may contain 1 to 3 heteroatoms selected from N, O and S, wherein the heterocyclyl group may be further substituted with one or more substituents selected from Ci-6 alkyl, OH, and halogen; R3 is selected from halogen and C1 -6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; is selected from 0 and 1; and n is selected from 0, 1, and 2; provided that Sm
R 1 is not 0 or OH when C is selected from phenyl and 6 membered monocyclic heterocyclic group and Ro is H.
13. The compound according to claim 12, or a stereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt, or co-crystal thereof, wherein R3
(R 2 Ro (R2 R is or Ro is selected from H, C1-4 alkyl, and cyclopropyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
R 1 is selected from 0, 0 o t Ria
D OH CN, CN, CN, 0 OH
.O 0 , and 0 Ria is selected from H, CI-6 alkyl, and -C(=O)C1.6 alkyl; R 2 is selected from cyano, -C(=O)NR2aR2b, C1-6 alkoxy, halogen, -S(=0)2R2a and
-C(=)OC 1 6 alkyl; wherein the alkyl or alkoxy is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C1 .6 alkyl and 3- to 5-membered cycloalkyl, wherein the C 1 .6 alkyl may be further substituted with one or more substituents selected from OH, D, and halogen; R2 a and R2btogether with the atom to which they are attached may form 5- to 6 membered heterocyclyl group which may contain 1 to 3 heteroatoms selected from N, O and S, wherein the heterocyclyl group may be further substituted with one or more substituents selected from OH and halogen; R3 is selected from halogen or C1 .6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is selected from 0 and 1; and n is selected from 0, 1, and 2; provided that m
Ri is not 0 or OH when C is selected from phenyl and 6 membered monocyclic heterocyclic group and Ro is H.
14. A compound of general formula (I"), or a stereoisomer, solvate, prodrug, metabolite, pharmaceutically acceptable salt, or co-crystal thereof, Ro
R3 NN O
(Rn C NH N \R1 (I",) wherein C is selected from phenyl and 6-membered monocyclic heterocyclic group, wherein the heterocyclic group may contain 1 to 3 heteroatoms selected from N; Ro is selected from H and C1 .6 alkyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
R 1 is selected from O, 0, O0, OH , CN,
and R2 is selected from cyano, carboxyl, -NR2aR2b, -C(=O)NR2aR2b, C 1.6 alkoxy, C1 .6 alkyl, halogen, -S(=0) 2 R 2 a and -C(=O)OC 16 alkyl; wherein the alkyl is optionally substituted with one or more substituents selected from halogen and deuterium;
R2a and R2b are selected from H, C1-6 alkyl and 3- to 5-membered cycloalkyl, wherein the C1 .6 alkyl is optionally further substituted with one or more substituents selected from OH, halogen, CI6 alkyl and C1 -6 alkoxy; R2 a and R2b together with the atom to which they are attached may form 5- to 6 membered heterocyclyl group which may contain 1 to 3 heteroatoms selected from N, o and S, wherein the heterocyclyl group may be further substituted with one or more substituents selected from C 1-6 alkyl and halogen; R3 is selected from halogen and C1 .6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is selected from 0 and 1; and n is selected from 1 and 2.
15. The compound according to claim 14, or a stereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt, or co-crystal thereof, wherein R3
R3 R3 (R c(R2 " N r R is selected from and Ro is selected from H and C 1 .4 alkyl, wherein the alkyl is optionally further substituted with one or more substituents selected from halogen and deuterium;
R 1 is selected from Ri is selected from 0 , 0
OH CN and R2 is selected from cyano, -C(=O)NR2aR2b, C1-6 alkoxy, halogen, -S(=0)2R2a and -C(=)OCi-6 alkyl; wherein the alkyl is optionally substituted with one or more substituents selected from halogen and deuterium; R2a and R2b are selected from H, C1-6 alkyl and 3- to 5-membered cycloalkyl, wherein the C1 .6 alkyl is optionally further substituted with one or more substituents selected from OH and halogen; R2 a and R2b together with the atom to which they are attached may form 6 membered heterocyclyl group; R3 is selected from halogen and C1 .6 alkyl, wherein the alkyl is optionally further substituted with 1 to 3 substituents selected from D and halogen; m is selected from 0 and 1; and n is selected from 1 and 2.
16. The compound according to claim 14 or 15, or a stereoisomer, solvate, metabolite, prodrug, pharmaceutically acceptable salt, or co-crystal thereof, wherein the compound is selected from
FC ~ N /' F >- NN N >zzzO NN N A- N N N -C N NCN H bH b 0 0 0 00 N N N NN fl ~ N- N H2,N N NN N N >=o 0 N> - NN H H H
0 0 0 / 0 /0/ NC, N 0N, "-= N N
N- NN A N>= N N>NN n' " N N>=z H H bHb 0 0 0
/10 n /-NN /= 1 N F N nk NN = N NN N N~ I CIN N N
b F H F H 0 0 0 0 F /NN, F /0/ HN N N NN NHN CIN NN NAN N HANN H bH bHb 0 0 0
0 //0/ N N NN NC NIN H2 N N N N H N :N>=o ~ NANN N - NN N H bH bHb 0 0 0
N N NN N ' NN N ~ 'NN NN N - NA >= 0 H N~A >z=o H N-A 'C> N NN N NN N N H bH bHNb 0 0 0 0 /0/ Nl N NN N NNN 0/ HN -' A = N H N K-A- N = > ,NNNN NN N N N b ~NH NN b 0 0
0 NCD CD3 N~
HNN N = "' N" N)= 3"N'
I I >zz0- N N "-- N N N.XH N
NC. N NCN. N NC-ONj- N>N N N 0 NC NN1 N== H. !"1NN/'= ~N N
-N N N N ,N ,N H N N N =HH H
0o , N CN OH,
NCON N N'C -a/-:>o N J N. N=71 N CF.3N NN/ NCN NN NH .N .N HNH I>= N N o H ,j N"N N N_ N N Y c0HO HO 00 0 0 0 N
N N NN.NH 2 N N ,JN N >1 =O H2N K\-' NN
N - N N F N N F tON H NN
N HOb 0CN ,CN,
0 H 2N N.1 I N o== 0 0 F N N H2N N. N NH 2N N FN NN F N~"
H H
0 0 0
N . N- N H2N -_ >=O0 ~ N .N.N N N N 0 F N "N N F N "N N H _ H H
' F N N N H
CN bNb
0 0 D3/, 3C N=1-, N/ N. N N DCN H2 N N. \
0H 0 0 0 -f / N.N NN N N~~ N NN N NN. H0~ b H b~ N. ~ N0 N Nj NN NH ~N N H~ H
N 0 N / I 0
N N NHNN N . N .
N N _N N>- N' N N N H ~N N HOb HOb H HO 0 0
D3CN /\N=N =0. N N H 2 N" N. NN N
NN NNN
H 0 , and H 0
17. Apharmaceutical composition, comprising: (1) acompound according to any one of claims14to 16, or astereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, (2) optionally one or more additional active ingredients; and
(3) a pharmaceutically acceptable carrier and/or excipient.
18. Use of the compound according to any one of claims 14 to 16, or stereoisomer, solvate, metabolite, pharmaceutically acceptable salt, co-crystal, or prodrug thereof, or the pharmaceutical composition according to claim 17, in the manufacture of DNA-PK inhibitor.
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